gwilliam@crc.ac.uk (Gary Williams x3294) (12/03/90)
The UK Human Genome Mapping Project would welcome contributions from the
Human Genome community to their quarterly newsletter G-NOME NEWS. The
copy deadline for inclusion in the next edition is 10th January, 1991.
Please send your articles to:
Dr. Nigel Spurr,
ICRF,
Clare Hall Laboratories,
Blanche Lane,
South Mimms,
Herts,
EN6 3LD.
Contributions can be accepted in any form - written, fax (0707 49527),
on disk (any format) or by e-mail (N_SPURR@UK.AC.ICRF)
========================================================================
G-NOME NEWS No. 4
------------------
The Newsletter of the UK Human Genome Mapping Project
Number 4 October 1990
Editors: Nigel K. Spurr
Enquiries to:
Nigel K. Spurr,
ICRF Clare Hall Laboratories,
Blanche Lane,
Potters Bar,
South Mimms,
Herts. EN6 3LD
Tel: 0707-44444, Ext. 353
Contents:
Page No.
1) Editorial - Nigel Spurr 3
2) UK Human Genome Mapping Project Resource Centre 4
a) Introduction to the Centre - Ross Sibson 4
b) Mapping the mouse genome in the HGMP 6
c) Human cell bank 6
d) HGMP Computing 7
e) UK DNA Probe Bank - Nigel Spurr 8
f) Random cell panel resource 9
3) PCR primers - Nigel Spurr & Sue Povey 11
a) New primers 11
b) Amendments to primers published in G-String 3 12
4) Human Genome research interests in Oxford - Ian Craig & 13
Veronica Buckle
5) A strategy for genome analysis - Hans Lehrach 23
6) Directed programme project grants awarded 26
(as at 1/2/90) - Furzana Bayri
7) Provision of expenses for attendance at HGMP meetings 30
8) User Registration for access to the services of the 31
Resource Centre - Christine Bates
1. Editorial
Nigel Spurr
Imperial Cancer Research Fund
Clare Hall Laboratories
Blanche Lane
South Mimms, Potters Bar, Herts. EN6 3LD
The publication of the fourth UK Human Genome Project Newsletter occurs
at a time when the MRC-supported UK Project now appears to be establishing
itself. The directed programme has supported a number of new projects and
enhanced equipment and resources in various participating laboratories (item 7
in this Newsletter details the proposals supported to date). Secondly, the
HGMP Resource Centre has now been equipped and staff are being trained. It
will not be long before the Resource Centre should be playing an important role
in the supply of materials to research groups. Though obviously it is hoped
that the Centre will play an interactive role and not be just a passive
supplier of resources.
The establishment of the Resource Centre coincides with the first of
the two Human Gene Mapping Workshops (HGM10.5 and 11) being held in Oxford and
London in September 1990 and August 1991 respectively. These meetings will be
the opportunity for the human gene map to be updated and to access the progress
of the various national Human Genome projects. It is anticipated that these
major workshops in Human Genetics will focussed international interest and will
be the ideal opportunity for these funding bodies to see how their money is
being spent in our understanding of human genetic diseases. The next
Newsletter will contain a summary of the meeting in Oxford taking place this
month.
The format of the Newsletter is now becoming established and it is
hoped that detailed articles on individual aspects of genome research will be
featured (item 6 by Hans Lehrach) alongside summaries of activities in the
various centres around the UK. This edition contains a summary of research in
Oxford compiled by Ian Craig and Veronica Buckle. I am still awaiting willing
volunteers to write short articles on their research or to help compile a
summary of work in their locality.
2. UK Human Genome Mapping Project Resource Centre
a) Introduction
Ross Sibson
HGMP Resource Centre
Clinical Research Centre
Watford Road
Harrow
Middlesex. HA1 3UJ
The UK HGMP differs from many other similar national endeavours in that
it has budgeted for a resource centre. Given that there are no plans for a
similar entity in other substantially better funded projects, why has the UK
adopted such a policy and where is it taking us? The answers to these questions
tell us what the Resource Centre is, what it is being set up to do and who
stands to benefit.
The origins of the Resource Centre lie in the needs of the UK human
genetics community to pursue a strategy for maintaining a credible
international position in the face of increasing competition. Despite the
difference of almost an order of magnitude between the UK and US budgets the UK
is already making a substantial contribution to the international effort to map
and sequence genomes. This is manifested in part by our strong representation
on international steering committees. Nevertheless, the UK scientific
approach, in common with that of its partners, is driven by the ambitions of
individuals. As yet the sum of the efforts of individuals remains an
intangible commodity. Even the most ambitious amongst us has not yet
undertaken to map the human genome in isolation. There is therefore a need for
a national approach which taps the substantial existing resources and gives
rise to measurable progress in mapping and sequencing genomes. The means to
distribute the ensuing benefits was also required. Furthermore, this approach
needs to enhance, not interfere with the flair and productivity of individuals
pursuing important related biological issues. Herein lies an inconsistency
because individual research groups cannot be expected to conform to a uniform
and systematic approach required to map and then sequence a genome. Nor would
they want to feed the postal service with responses to the enormous number of
requests likely to be attracted by their activities.
These problems faced the MRC when they were appointed custodians of the
additional funding already earmarked by the government for an extra initiative
in genome research. In particular, how could they produce something tangible
to show following their period as trustees. The solution had two components.
One was a directed programme steered by a committee which aimed to allocate
funds where they could be used immediately to accelerate genome research. The
second was a Resource Centre which would act as a focus for collection and
distribution of important genetic materials. This would ensure as efficient
access as possible for the whole of the community. The Resource Centre would
also perform the bulk of the procedures required to sustain a systematic
programme. It would therefore need a focused group whose activities would free
researchers to concentrate on the biological issues which emerged.
The Resource Centre now has a physical location in the CRC at Harrow
but this is just one manifestation of what is likely to remain a distributed
resource. Other manifestations include the collection of data on cytogenetic
abnormalities run by Margaret Fitchett of the Oxford and Pat Jacobs of the
Wessex Regional Cytogenetic Laboratories, respectively and will be available
through a Human Cell Bank at Porton run by Alan Doyle and Bryan Bolton as part
of the ECACC.
It has turned out to be surprising how few UK groups have ready access
to good quality sequence manipulation software and databases. This will soon
be corrected when major packages become available from the resource centre to
the whole of the UK through JANET. Packages for manipulation will include the
Staden software and GCG (the University of Wisconsin Genetics Computer Group
package), genetic linkage packages such as Latharop and Lalouel's and
Felsenstein's Phylogeny Inference Package (Phylip), while databases on line
will include GENBANK, EMBL, NBRF, SWISSPROT, VECBASE, OMIM and GDB. There will
also be a range of other useful packages including Electronic Mail and BIOSCI,
the Biologists Bulletin Boards from which information as diverse as contents of
biological journals to Scientific interest groups can be obtained. Computing
groups at the CRC and Cambridge have been instrumental in getting the system in
operation. Additional hardware has had to be installed. The main items are a
distributed array processor which will greatly enhance, for example, exhaustive
sequence searches and a file server. These are presently located at Cambridge
and linked to the CRC via a land line. Prospective users of the network may be
relieved to know that great efforts have been made to make it as user friendly
as possible and that these efforts will continue in the future. It is intended
that the network will be used first by a few trialists and will then go on
general release before the end of the year. The biological and the computing
activities of the resource centre will include training courses for interested
parties.
The Resource Centre with walls (as it has been called) is now underway
in the CRC at Harrow. A building has been refurbished (to include abundant
access), equipment installed and staff recruited. The staff are now busily
training for this new role. Part of their training is performed under the
guidance of key UK academics like those in the group of Sidney Brenner at
Cambridge. The aim is to introduce services as soon as they become available
and to have a full service by the end of February 1991. Our first offering is
likely to be the ICRF probe bank, managed by Nigel Spurr. Arrangements have
been made to transfer the bank under Nigel's guidance to the Resource Centre
where it will be available this autumn. The Probe Bank will be further
developed at the Resource Centre and will also include oligonucleotide primers.
No reference has been made so far as to what the systematic work will
entail. It certainly will not be the short arm of a not very interesting
chromosome and will include sequencing. In fact, the formally agreed UK
strategy is to concentrate our limited resources on sequencing new cDNAs and
locating them on physical maps of the genome. Ideally, priority will be given
to those selected or provided by users. This has a number of other important
advantages besides being within our resource constraints. It can use existing
procedures to generate important information in a short space of time. New
cDNAs can be used as unique probes for mapping purposes. This has the added
advantage that the corresponding genomic sequences will be spread over a larger
part of the genome than is absolutely necessary to code for the cDNAs. The
cDNA strategy also adds knowledge incrementally thus sustaining the progress of
the directed program.
Inevitably, the decision to pursue a particular strategy raises more
issues than it solves. This is particularly true when one transfers it to an
operational level. A steering committee presently comprising Bob Williamson
from St. Marys, George Stark from the ICRF and Sidney Brenner from Cambridge
has therefore been set up to oversee the activities of the Resource Centre.
They will meet on a regular basis to ensure that the direction is consistent
with the needs of the whole of the community. In addition, they will be aided
by ad hoc working parties selected from individuals active in key areas to
address issues relevant to those areas. The working parties will largely be
concerned with a technical agenda. This is illustrated by the example of
screening YAC libraries. The groups of David Bentley at Guys and Kay Davies at
Oxford have acted very efficiently to transfer the original Olsen and
Schlessinger libraries from St. Louis and are now developing the best
screening methods. These libraries are earmarked for the Resource Centre.
Recently, other libraries have become available. We are therefore in an
enviable position but how can it be used to everyones advantage? Should we
screen every clone? If not which ones? Should the screening be performed at one
centre or several centres? The answers to these questions rely heavily on
technical issues. Is it practical to distribute the libraries? Should they be
distributed in a form which is suitable for screening?
The core group also has to get to grips with non-technical issues which
nevertheless have a bearing on the science. Who will want YACs corresponding
to their probes and on receipt of the YACs will their problems be just
beginning? This raises the wider issues of what happens to the data and who is
the constituency for the Resource Centre? Once the constituency has been
defined it becomes easier to determine its needs. These needs turn out to be
very diverse because the users also form a diverse group. The group is likely
to range from clinicians with a particular patient group who need to gain
access to a repertoire of molecular genetic techniques and materials, to
evolutionists gripped by the implications of each new cDNA as its sequence
rolls out of the production line, front line molecular biologists aiming to
gain insight into the whole shooting match by perturbing expression in vivo
through the delivery of novel constructs. Even probe collectors are expected.
The constituency must also include industrialists. One of the
conditions attached to the extra funding is that it should be used in a way
that can benefit UK industry. Here the issue of access to data may be at its
most important. The expectation is that users of the Resource Centre should
also contribute to it. One obvious way is that the information obtained when a
user supplied probe is used to screen for example a gridded YAC library should
contribute to the development of a physical map based upon the library.
Information about the interests of say a pharmaceutical company could
potentially be obtained by screening the data base for their contributions.
This is where a Resource Centre comes into its own because it is impartial and
can introduce appropriate safeguards into a database. A lag before information
is released could for example be easily introduced. We may have here a
seemingly illogical concept which could serve well, that of a private public
database.
I look forward to the palmy days ahead when all 100,000 cDNAs have been
sequenced and 100% of the genome is available in fragments that have been
physically mapped. There is an unfortunate statistic about the early
operations of the Resource Centre. Disappointment is guaranteed for a certain
percentage of its users and the number of disappointments will be in direct
proportion to the use made. Prospective users should not feel discouraged.
Catalogues will soon be available with listings of resources such as probes and
primers and these will be maintained on a supply and demand basis. It is where
results cannot be guaranteed, especially in the case of screening, where the
risk of disappointment will be greatest. This is the main element of the
service which will develop through interaction with the community. Those who
remain in doubt should consider, given the high level from which the original
seal of approval came and the increasing need to sustain domestic capability,
whether this is an opportunity that can be wasted.
b) Mapping the mouse genome in the HGMP
In their early discussions, the Directed Programme Committee agreed to
focus on mouse genome mapping as one element in their strategy. Several grants
were awarded (see the listing of grants in this number of G-String). To
sharpen up this focus, the DPC sponsored a meeting, held in London on 6th April
of this year and organised by Dr. Steve Brown (St. Mary's), of
representatives of the mouse-genome-mapping community - including teams in
mainland Europe and the US. A report went to the July meeting of the DPC and
its recommendations, on opportunities, needs and priorities, have been broadly
accepted as a basis for HGMP policy in this area.
COPIES OF THE REPORT ARE AVAILABLE (free-of-charge) from the HGMP
Resource Centre; telephone Joanne Grewcock on 081-869 3808.
At the Users Meeting on 30th April, Dr. Philip Avner from the Pasteur
Institute gave a guest lecture on the relevance of mouse-genome-mapping to the
analysis of the human genome.
Following this policy formulation, the Project Management Committee
agreed to set up a mouse-genome-mapping resource at the Resource Centre, in
collaboration with the Pasteur Institute and the Comparative Biology Section at
the MRC's Clinical Research Centre.
DNA is being prepared from a large-scale backcross between Mus musculus
and Mus spretus, and in the first phase of the programme a consortium of major
groups will collaborate in generating a global anchored STS map of the genome.
Grants are being negotiated to make mouse cDNAs for use as probes. It is
planned to offer, from early 1992, a rapid mapping service for users. In
support, a genetic mapping database and mouse probe reference library will be
set up.
As an immediate service, GBASE (The Jackson Laboratories mouse
database) is available on- line from the Resource Centre. We are grateful to
the Jackson Labs for their generosity in providing this facility and for
permitting their Manual to be reproduced: copies are available from the
Resource Centre (see above). Support for the Harwell Group has been provided,
to get their data similarly available on-line.
c) Human cell bank
The Centre for Applied Microbiology and Research at Porton now operates
a Human Cell Bank as part of the European Collection of Animal Cell Cultures
(ECACC). The HGMP funds user-access to this Bank in much the same way as the
Research Councils jointly provide block-grant funding for access to the parent
Collection. Catalogues and details of services are available on request from
the Bank (and will be sent to those who signal their interest on the
Registration forms):
PHLS Centre for Applied Microbiology and Research Porton Down Salisbury Wilts.
SP4 0JG
Tel: 0980 610391 Ext 511. Fax: 0980 611315
To have access to the Bank's services free of cost, you will need to
register as a Resource Centre 'user'. The price is the usual quid pro quo: it
will have to be freely available to other registered users. It will still be
possible to place restriction on access to the line by non-HGMP customers of
the Bank.
Major uses of the Bank's services will not be automatically covered by
the HGMP funding arrangements; for these, funding will have to be found from
other sources. For the time being, commonsense will be a guide as to what is
"major", but in cases of doubt discuss your requirements at the planning stage
with Bryan Bolton at the Bank. It may prove necessary, in due course and in
the light of experience, to impose restrictions to ensure that the Bank does
not fill up with cell-lines that are irrelevant to the objectives of the HGMP.
A Liaison Committee has been set up to keep such questions under review, to
define accession policy and encourage the deposit of valuable cell-lines, and
to monitor HGMP use of the Bank, especially in these early stages. Any
comments or suggestions from users should be directed to the Secretary of the
Liaison Committee, Dr. Furzana Bayri, at MRC Headquarters (071-636 5422).
d) HGMP computing
Those who can use the Joint Academic Network (JANET) should now be able
to access the HGMP Resource Centre's computing facilities once they have
registered as a 'user' and been issued with a user id and password. At this
stage we simply do not know how much traffic there will be. Until it becomes
clear that we are running close to the limits of existing computing capacity,
it seems sensible to let anyone who wants access to have it, irrespective of
their interests. That means that the criteria for registering as a computer
user will, for the time being, be more relaxed than for those who want access
to the other services (including computer training courses) we are offering.
A substantial proportion of prospective users do not have access to
JANET. All those on academic sites should be able to be linked; we will do
what we can to help, but the first initiative must be local: identify the
person who is responsible for the network operation on-site and try to get
things sorted out.
Until now, there have been two registration forms: one, for computer
users, pre-dated the formal setting up of the Resource Centre, and was used
mainly for those attending the first computer training courses. Those who
registered using that form will continue to be registered for computing access
only. If they want access to the wider range of facilities, then they must
fill in the later form (available from Joanne Grewcock at the Resource Center:
telephone 081-869 3808). In future, all users should fill in the later form;
if they indicate interest only in computing access, then the relaxed criteria
for registration will apply, at least until we begin to reach the limits of
computing capacity.
There are already signs of a misapprehension that access can be on an
easy-going departmental or group basis. That is not the case. Each individual
user must register and have a personal password. Passwords must be kept
secret. Even if we were stupid enough to discount the damage that 'hackers'
might do to our systems, we would still be dependent on access to other systems
that do require precautions.
To begin with, the system will be operating as a beta-test, to get it
working efficiently. Users in these early days will have to understand that
there may be problems. It will be most helpful if they can be reported to us
on the help line (number in the computing manual). The fact that we have got
to this stage so quickly is largely due to the efforts of Francis Rysavy and
his colleagues of the Clinical Research Centre's Computing Services Group.
We shall continue to depend heavily on the Computing Services Group
expertise, but Dr. Martin Bishop - who has already been closely involved in
the computing side of the HGMP - is now appointed as the Computing Manager of
the Resource Centre. He will divide his time between the Molecular Genetics
Unit at Cambridge (0223 402406) and the Resource Centre.
The Resource Centre computing facilities already include access to most
of the standard databases and software packages relevant to genetics. We are
currently in discussions with the Howard Hughes Medical Institute and Johns
Hopkins University with the objective of running from the Resource Centre the
new Genetics Database (GDB) - launched successfully at the recent HGM Workshop
10.5 in Oxford - to provide a UK node for both read-only access and editing
access. Two parts of the package - the Jackson Labs Mouse database (GBASE) and
Victor McKusick's OMIM - are already accessible on a trial basis. Contact
Joanne Grewcock for user manuals for these packages. We are extremely grateful
to all the protagonists in the US for being so helpful and generous. Again, we
should warn that running the new GDB is not a trivial exercise, and we shall
have to sort out problems as we go.
There will be an immediate need for training in the use of the new
packages, and that will be the first priority in scheduling of training courses
for the rest of this year).
e) UK DNA Probe Bank
Nigel Spurr
Imperial Cancer Research Fund
Clare Hall Laboratories
Blanche Lane
South Mimms, Potters Bar, Herts. EN6 3LD
Once a person has registered to use the HGMP resource centre they will
(within reason) be able to obtain DNA probes upon request. There are currently
650 DNA probes available in our catalogue, the majority of which have been
assigned to chromosomes and detect RFLPs. A simplified listing of probes and a
more comprehensive catalogue is now available. If you want a copy please
contact Christine Bates at the HGMP Resource Centre:
HGMP Resource Centre
Clinical Research Centre
Watford Road
Harrow
Middx. HA1 3UJ
Tel.No: 081-869 3446
Fax No: 081-869 3807
DNA probes will be distributed as purified DNA usually in 5 mg aliquots
in 10mM Tris, 1mM EDTA pH 7.4. The majority of the DNA has been prepared using
the Quiagen separation system and all samples are quality controlled by
visualisation on agarose gels after restriction enzyme digests to excise
inserts. If larger quantities of DNA or bacterial stabs containing
recombinants are required please contact the Centre to discuss your
requirements. These will only be available in exceptional cases.
After the 17th September 1990 all DNA probe distribution will occur
from the Resource Centre, therefore all requests for probes should be sent to
the Centre for processing. There is an order form (and a release agreement) in
the catalogue and from now on no probes wiill be issued except on the receipt
of the standard form. No material will be distributed without a written
request though telephone calls to discuss particular needs etc. will still be
welcome. Some probes have particular release conditions and a user will be
made aware of these at the time of request.
It is anticipated that interactions with the UK DNA Probe Bank will be
two-way and not a distribution service only. If you have probes you would like
to include in the Bank we would be happy to include these in our listings.
Similarly, if you have specific requests for probes not currently held, we will
try and ensure that we obtain these.
One of our aims is to clone new markers particularly for regions of the
genome at present under-represented. Therefore if you have specific ideas or
collections of unsorted probes that may be useful in this endeavour please
contact me at the UK HGMP Resource Centre or at the ICRF Laboratories at Clare
Hall.
f) Random cell panel resource
Several laboratories have expressed a requirement for a random control
panel in order to define normal UK frequencies of genetic markers. Such a
panel would be of use in epidemiological and disease studies as well as studies
of newly identified genetic polymorphisms.
We are currently investigating the possibility of such a resource being
set up at the Public Health Laboratory at Porton Down which already houses the
European Collection of Animal Cell Cultures.
In order to ensure the unbiased nature of the selection of cells to be
used it is proposed that the cell panel should in the first instance consist of
cells from cadaver kidney donors, collected from all parts of the UK over the
last few years. A panel of about 240 cells from these donors collected by the
National Tissue Typing Research Laboratory in Bristol is available for
transformation into cell lines. It is still to be decided how many of these
will be transformed for the first random control panel.
Funding for this project is being sought from the Medical Research
Council and the size of the panel and service provided for its use will to a
large extend depend on the expected use that will be made of the panel.
Therefore, if you think that your laboratory would make use of such a resource,
it would be helpful if you could fill in the attached note and return it as
soon as possible to Dr. Nigel K. Spurr, ICRF Clare Hall Laboratories, Blanche
Lane, Potters Bar, South Mimms, Herts. EN6 3LD.
Name of Investigator:
Address:
1. I would use a random cell panel if available Yes No
2. I would prefer to receive A. Viable cell lines
B. DNA samples
3. I would expect to request samples A. Once a year
B. More often
C. Less often
4. For my studies a minimum panel size
would be A. 100 cells
B. 200 cells
C. State number
5. I would be prepared to return data on the genetic markers tested to the
cell databank as a condition of receiving the cell lines or DNA.
Signed: ...............................................................
Date: ...............................................................
Please return to: Dr. Nigel K. Spurr, ICRF Clare Hall Laboratories, Blanche
Lane, Potters Bar, South Mimms, Herts. EN6 3LD
3. PCR Primers
Nigel Spurr
Imperial Cancer Research Fund
Clare Hall Laboratories
Blanche Lane
South Mimms, Potters Bar, Herts. EN6 3LD
Sue Povey
MRC Human Biochemical Genetics Unit
The Galton Laboratory
University College London
Wolfson House
4, Stephenson Way
London NW1 2HE
a) New primer:
Name (sequence identified):
PENK, human proenkephalin gene
Primer Sequence: 1 5'-TAATAAAGGAGCCAGCTATG-3'
2 5'-ACATCTGATGTAAATGCAAGT-3'
Chromosomal location:
8q23-q24
Conditions of use:
94oC 1 min 0-5-1mg DNA
55oC 1 min 300ng each primer Total reaction
72oC 1 min 10mM triphosphates volume of 100ml.
25-30 cycles
Detection system:
On 10% neutral acrylamide gel or 2% agarose
Use:
Linkage analysis 79bp repeat fragment
Reference:
Weber, J.L. and May, P.E. (1990) Nucl. Acids Res. 18, 2200.
b) Amendments to primers published in G-String 3:
1. PGAM2 ) Need at least 30 cycles and
C9 ) triphosphate final concentration
P1 (a1 antitrypsin) ) should be 1.5mM not 15mM.
2. PGAM2 detects a 334bp fragment.
3. C9 only needs a 10 second first step at 90oC.
4. ALDOB. A different set of primers have been described:
Primer sequence: 1 5'-TCATTGCTTGCTTTCTCAAGCAGGG-3'
2 5'-CAATGCTTCTCCGTGTTGGAAAGTC-3'
Reference: Tolan, D. and Penhoet, E. (1983) Characterization of the
human aldolase B gene. Mol. Biol. Med. 3, 245-264.
We would like to thank Cathy Abbot for her assistance and the corrections
particularly to the ALDOB primers.
4. Human Genome Research Interests in Oxford
Ian Craig and Veronica Buckle
Genetics Laboratory
Department of Biochemistry
University of Oxford
South Parks Road
Oxford OX1 3QU
Background to the development of human genome interests at Oxford
On the 13th February 1965, Watkins and Harris reported (from the
University Pathology Department) that inactivated Sendai virus could enhance
fusion between animal cells from different species, thereby opening the pathway
to routine preparation of human-rodent hybrids. The first panel of somatic
cell hybrids for human gene mapping in the UK was brought to Oxford in 1970 by
Walter Bodmer, who had been appointed to the newly created Chair in Genetics
associated with the University Biochemistry Department, and his research group
including Markus Nabholz. Nearly a decade of mapping activity under Walter
Bodmer's direction in the Genetics Laboratory followed; however, the emphasis
was always on biological significance and mapping was employed as an analytical
tool in solving wider problems. The Genetics Laboratory has had a participant
at all Gene Mapping Workshops including the first in Yale, HGM1, 1973. Walter
and Julia Bodmer were at that time particularly interested in the genetics of
the HLA system. Many of those training under Walter Bodmer in Oxford have
continued to make major contributions to human genetics. Those remaining in
the UK include Veronica van Heyningen (Edinburgh), Peter Goodfellow, Ellen
Solomon and John Trowsdale (all now at the the ICRF) and Jonathon Wolfe (U.C.
London), who was a fly geneticist in the Genetics Laboratory, and was obviously
seduced by the allure of higher organisms. The development of techniques for
the analysis of chromosomes in patients and in hybrid cell lines was strongly
influenced by Martin Bobrow (now Professor of Paediatrics at Guy's Hospital)
who was associated with the Genetics Laboratory during the 1970s.
Martin Bobrow and Peter Pearson had previously been in the MRC
Population Genetics Unit at Oxford (Director, Dr Alan Stevenson). Another
notable worker associated with the early days of the Genetics Laboratory was
Alec Jeffreys who, as a graduate student in Ian Craig's laboratory, exploited
the segregation of human mitochondrial DNA in most rodent human hybrids in the
analysis of mitochondrial gene products. Others in the group later provided
the first characterisation at the sequence level of mitochondrial DNA mutations
(conferring chloramphenicol resistance). Alec Jeffreys was later (1979) to
publish the localisation of the beta globin locus to 11p with members of the
Genetics Laboratory; this was the first gene assignment by Southern blot
analysis of somatic cell hybrids. Although the hybrid legacy remains with the
Genetics Laboratory and has been applied to a variety of studies, including the
detailed characterisation of chromosomal translocations in girls with Duchenne
muscular dystrophy, such approaches have been augmented on the one hand by the
resurgence of family studies and on the other by development of physical
mapping techniques.
Analysis and clinical applications of recombination have been guided by
Professor John Edwards who took over the Chair of Genetics from Walter Bodmer
in 1979.
Physical mapping has been supported through development of the
"waltzer" PFGE approach under Professor Edwin Southern (Biochemistry
Department), whose other contributions to the development of genome research
are known to all in the field. In situ hybridisation of single copy sequences
is another physical approach which was integrated into human genome research at
a relatively early stage; collaboration between Pathology and Genetics
resulting in the subregional localisation of Factor IX in 1983.
Professor Rod Porter's MRC Immunochemistry Unit in the Biochemistry
Department specialised in protein isolation and characterisation of
immunoglobulins, complement proteins and other members of the major
histocompatibility complex. Before his untimely death, he anticipated the
contributions of molecular biology and set the course of the Unit firmly in the
direction of analysis at the DNA level, an approach which it has continued to
exploit to good effect.
In the Sir William Dunn School of Pathology, interests in somatic cell
genetics continued under Professor Henry Harris through the analysis of
malignancy in hybrids. The first irradiation fragment hybrids were
characterised there by Stephen Goss. Yvonne Boyd (one of Henry Harris's
graduate students, now at the MRC Radiobiology Unit) worked on chick/rodent
hybrids before becoming involved in human gene mapping in Edinburgh (MRC CAPCU)
and then, returning to Oxford, applied her expertise to the analysis of DMD
translocations. Christopher Marshall was a graduate student at the same time
and now works with oncogenes at the Chester Beatty Institute. Peter Cook's
group have made important progress in the investigation of chromatin structure
and nuclear organisation. The arrival, in 1980, of Professor George Brownlee
brought DNA expertise to studies on influenza virus and blood clotting factors.
These remain the main focus of the group and the cloning of the Factor IX gene
was one of the first successes concerning the isolation of genes whose products
have highly significant pharmaceutical implications.
Interests in the haemaglobinopathies at the John Radcliffe Hospital
flourished under Professor Sir David Weatherall and the MRC Unit of Molecular
Haematology was founded in 1981. Subsequently, under his guidance and in
collaboration with other clinical research interests, the Institute of
Molecular Medicine (IMM) was established in 1989, a venture sponsored jointly
by the MRC, the Wellcome Trust, the ICRF and several other charitable
organisations together with the University of Oxford. The Institute has been
set up so that its work is integrated into the different clinical departments
in Oxford. A considerable amount of the work in the Institute is related to
genetic disorders which is reflected by collaborative ties with the Department
of Medical Genetics at the Churchill Hospital, and many of the groups within
the Institute have links with the basic science departments in Oxford.
Over an equivalent period, key discoveries with far reaching
implications for mammalian genetics concerning X-inactivation, sex
determination and genomic imprinting were made in the Genetics Division of the
MRC Radiobiology Unit at Harwell, using mice as the model system (M.F. Lyon,
B.M. Cattanach and A.G. Searle). The repercussions of these discoveries have
continued effects in the studies of several groups who are involved in the
analysis, of mouse mutations, including those of interest as potential
homologues of human syndromes, and as a route to investigating chromosome
inactivation. Recently, molecular approaches have been introduced both for
genetic analysis in mice and for physical and recombination mapping in humans.
Current Human Genome research in Oxford and its environs is thus spread
widely and covers a range of technologies from the development of new
approaches to physical mapping and sequence analysis to detailed studies on
gene function and expression. Areas which provide a focus for the research
activities of several groups include the analysis of genetic phenomena such as
inactivation and imprinting, studies of chromosome function and the control of
gene expression. The identification and analysis of mutations (at a functional
level in clinical defects) and the development of somatic gene therapy provide
topics for other collaborative work.
The breadth of contemporary human genome related research topics can
best be represented by the edited summaries of the various individual groups.
Any errors of omission, or commission, are the responsibility of the present
authors.
1. Institute of Molecular Medicine, John Radcliffe Hospital, Headington,
Oxford OX3 9DU.
[This includes the National Haemoglobin Reference Laboratory, which under John
Old's supervision provides a clinical and laboratory referral service for
haemoglobin disorders throughout Britain. It also acts as a training centre
for clinicians and scientists wishing to develop a prenatal screening
programme.]
Douglas Higgs' Laboratory
We have established that the human alpha-globin complex lies within
approximately 200kb of the telomere of the short arm of chromosome 16 and is
associated with a group of constitutively expressed genes in this terminal
segment. Characterisation of naturally occurring mutations of this region,
which cause alpha-thalassaemia, has helped to identify the important cis-acting
sequences controlling alpha globin gene expression and further experimental
studies are in progress with transgenic mice and stable transformants in
erythroid cells. This combined approach is enabling us to characterise all of
the the cis-acting sequences that are required to regulate the tissue and
developmental-stage specific expression of the alpha-like genes. Our work
includes the analysis of a group of much larger deletions from the tip of
chromosome 16 that are associated with dismorphism and mental handicap in
addition to alpha-thalassaemia. Ultimately, we hope to identify which of the
many genes surrounding the alpha globin complex may be responsible for
producing the associated abnormalities. We have now completed a physical map
encompassing the entire region from the alpha globin genes to the telomere of
chromosome 16. This, together with a large number of naturally occurring
mutations that we have identified in this region, provides a most interesting
and informative model system for the analysis of human genetic diseases.
John Clegg's Laboratory
We are interested in the evolution and genetics of equid haemoglobins.
Horses have one of the richest fossil records of all mammalian species. Their
evolution can be traced in detail from 50 million years ago, when the line
split from the ancestors of rhinoceroses and tapirs, finally to give rise 5
million years ago to the forerunners of the modern equid family. The timing of
this last radiation closely parallels that of man, gorilla and chimpanzee, and
offers a valuable opportunity for comparative studies on the molecular
evolution of various gene loci. We are undertaking detailed comparison of the
alpha globin gene complex of equids and humans and determining the chromosomal
location of the equine alpha complex.
Bill Wood's Laboratory
We are studying the developmental regulation of the beta globin gene
cluster, particularly the switch from foetal gamma gene to adult beta gene
transcription which occurs in the perinatal period. This involves
investigation of natural mutations resulting in hereditary persistence of fetal
haemoglobin (HPFH); the expression of cloned fragments of the cluster and its
upstream regulatory region in mouse erythroleukaemia cells; and the use of
transgenic mice for studying changes during normal development.
Swee Lay Thein's Laboratory
Analysis of the molecular basis for the beta thalassaemias in India,
Thailand, and Burma including the genetic study of conditions in which there is
a defect in the switch from foetal to adult haemoglobin production. We are
characterising the modifying factors which affect the clinical manifestations
of beta thalassaemia with a view to improving prognosis and genetic counselling
for this disorder. We are also trying to localise the gene(s) responsible for
the trans-acting factors involved in HPFH in which the high F phenotype
segregates independently from the beta-globin cluster.
Peter Harris's Laboratory
We are involved in the construction of a long range map of the
chromosome 16 short arm between the telomere and the locus for polycystic
kidney disease with the ultimate aim of identifying and cloning the defective
gene involved in this disorder.
Veronica Buckle's Laboratory
The work of the cytogenetics laboratory is closely integrated with
other groups in the IMM and elsewhere in the University. We are currently
developing high resolution approaches to in situ hybridisation using interphase
nuclei, non-isotopic multi-labelling techniques and confocal microscopy which
will be of enormous help in the rapid assignment of sequences. Our studies on
the distribution of sub-telomeric sequences and the definition of
rearrangements not detectable by standard cytogenetic techniques are
contributing to an understanding of the nature of homologous pairing and
sub-microscopic telomeric translocations.
As part of the LRF Preleukaemia Unit at Oxford we are collaborating in
a molecular characterisation of the breakpoints associated with deletions
involving chromosome 5 in myelodysplasia and have demonstrated homozygous loss
of the CSF1 receptor in a subset of patients with chromosome 5 deletions. We
are also constructing a long range map of regions of chromosome 3 long arm as
part of a study of rearrangements of this chromosome associated with leukaemia.
Kay Davies' Laboratory
We are studying the molecular basis of several human genetic disorders
for which the biochemical defect is unknown. Considerable advances have been
made in studies on Duchenne (and the less severe, Becker) muscular dystrophies.
Over the last three years, we and others have cloned and sequenced the gene
involved. By analysing patient DNA our group has shown that the vast majority
of patients suffer from DMD or BMD because they have lost part of the gene
coding for dystrophin. This information has provided a means of direct carrier
detection and prenatal diagnosis. Current research efforts are concentrating
on identifying the function of dystrophin in the muscle cell in order to
understand the disease process. This is the next step in developing treatment
and potential cures for these diseases in the future. We have recently
characterised a protein that is very like dystrophin and we are investigating
its expression in DMD and BMD patients as well as in other muscular dystrophies
such as limb girdle dystrophy.
The fragile X syndrome is the most common genetic cause of mental
retardation after Down's syndrome, affecting approximately 1 in 1000 males and
is associated with fragility of the X chromosome at Xq27.3. We have recently
micro-dissected this region of the X chromosome and are currently isolating
sequences that correspond to genes expressed in brain.
Retinitis pigmentosa is a progressive eye disease that in some cases is
also due to a defect in an X-linked gene. Other workers have established where
the gene must lie on the chromosome and our research is now concentrating on
identifying candidate genes in this region. We are using our expertise with
libraries produced through microdissection of the X chromosome to isolate
candidate sequences that might be expressed in the retina.
The spinal muscular atrophies (SMA) as a group comprise the second most
common neuromuscular diseases of childhood after the muscular dystrophies.
Manifestation is variable, but, in its severest form, SMA has a very early
onset, sometimes in utero. Together with our clinical colleagues, we are
currently collecting families affected by SMA. Our linkage analysis has
localised the defect to chromosome 5 and we are now intensively mapping this
area as the first step in localising the gene defect. Both microdissection and
YAC technologies are being applied to this search.
As part of the Human Genome Mapping Project, one of the duplicate
copies of the St Louis YAC library is being transferred to the IMM, where it
will form the basis for a screening programme available to the UK research
community. This will be ultimately transferred to the MRC Resource Centre at
Northwick Park Hospital.
2. The Nuffield Department of Surgery, John Radcliffe Hospital, Headington,
Oxford OX3 9DU
Dr. John Todd
Analysis of the genetic basis of a complex, multifactorial disease: autoimmune
type 1 diabetes
We are developing methods and approaches to map susceptibility genes
responsible for the polygenic disease, Type 1 diabetes in humans. We have
collected over 100 multiplex families with at least two affected children in
each pedigree. We are currently using highly polymorphic (PIC > 0.7) VNTR-type
probes (both minisatellites and microsatellites using blotting and PCR methods)
to obtain linkage to disease. One strategy we have employed is to use the
mouse model of Type 1 diabetes to exploit the power of genetics in the mouse.
We have constructed a simple map of the mouse genome using microsatellites that
show variation between the diabetic strain and the diabetes- resistant strain.
With about 60 microsatellite markers we have detected significant linkage to at
least four genes that affect susceptibility to Type 1 diabetes in a backcross
pedigree of 92 animals. Fine mapping has revealed that one of the genes lies
within a known area of synteny between the mouse and human genomes. It is
hoped that since the murine and human diseases are very similar with respect to
autoimmune pathogenesis and genetics that this strategy will help
identification of human disease genes.
3. Molecular Immunology Group
John Bell's Laboratory
Genes which control the structure of the histocompatibility complex
(MHC), have been associated with susceptibility to more than forty diseases.
We have shown that the risk of developing diabetes mellitus, rheumatoid
arthritis and coeliac disease is determined by the type of HLA gene which an
individual inherits and we are mapping disease susceptibility loci for these
disorders. We are cloning lymphoid cDNAs by eukaryotic expression and have
begun to generate PCR based polymorphic markers on chromosomes 7,11 and 14.
4. Molecular Infectious Diseases Group
Mark Gardiner's Group
Mitochondrial disorders. Diseases caused by mutations in mitochondrial DNA
include mitochondrial myopathy (muscle weakness, mental retardation and fits)
and Leber's hereditary optic neuropathy (causing blindness). We have
identified two novel mutations and are studying the way in which they cause
disease. At present mitochondrial myopathy can only be diagnosed by muscle
biopsy, so we are developing a way of diagnosing them and detecting carriers on
the basis of a single blood test. We anticipate that this will provide a
stronger scientific basis for genetic counselling.
Batten's disease. Among neurodegenerative disorders, Batten's disease is
extremely rare but its study is illustrative of how modern genetic analysis can
be used to identify a genetic defect responsible for the accumulation of
abnormal macromolecules in the nervous system during childhood and their
interference with development and function. This project has used the
phenotypic and genotypic information made available from studying several
families affected by Batten's disease and has allowed the group to make a
provisional assignment of the defective gene to the long arm of chromosome 16.
5. Neurosciences Research Group
John Newsom-Davis's Laboratory
We are investigating the susceptibility of the neuromuscular junction
to autoimmune disease. We have cloned the genes for five acetylcholine
receptor (AChR) subunits and have mapped four of them (to chromosomes 2 and
17). Using recombinant AChR alpha subunit preparations of the complex produced
by expression vectors we are raising T cell lines and clones from Myasthenia
Gravis patients. These are being tested with smaller recombinant fragments and
with synthetic peptides to define the exact sequences of the AChR 'seen' by the
T cells. Mutations in the AChR genes of patients with congenital forms of MG
are being defined using PCR. Our research has shown that autoimmune mechanisms
also underlie the Lambert-Eaton Myasthenic Syndrome and, as part of this study,
we are in the process of cloning genes for calcium channel subunits.
6. Collagen Genetics Group
Bryan Sykes' Laboratory
Natural variation in the form of restriction fragment length
polymorphisms (RFLPs) has been used to follow the inheritance of collagen genes
in families with diseases which are likely to be caused by defects in their
collagen. Type 1 Collagen is coded by two genes. Our Group has shown that the
dominant form of Brittle Bone Disease, Osteogenesis Imperfecta is caused by
mutations in one or other of these genes. We now wish to pinpoint the
mutations, which are likely to be different in each family. Type 2 collagen is
the major structural component of cartilage. As such it is highly likely to be
involved in Generalised Osteoarthritis (GOA) which is characterised by a
breakdown in the cartilage. GOA does not usually develop until the age of 40
plus, so the use of multigeneration families is limited therefore a sib pair
analytical approach is being employed instead.
7. ICRF Cancer Research Unit, Institute of Molecular Medicine, John Radcliffe
Hospital, Headington, Oxford OX3 9DU
Molecular Oncology Group (Adrian Harris, Ian Hickson)
We are adopting various strategies to clone DNA repair genes from human
and other mammalian sources. One approach is the direct complementation of the
defect in patients with repair-deficiency syndromes such as Ataxia
Telangiectasia and in a range of laboratory induced hamster cell mutants.
Other strategies include the use of PCR to clone conserved domains identified
in bacterial and yeast repair genes, the isolation and sequencing of repair
enzymes and the isolation of DNA damage- inducible genes. We are currently
characterising two genes; an X-ray inducible gene that maps to 1p12-34 and one
encoding an enzyme that repairs oxidative DNA damage (not mapped).
We have cloned the human gene for topoisomerase II (17q21-q22) and are
studying regulation of expression under different conditions such as cell cycle
transit and growth factor stimulation.
Dave Simmon's Laboratory
We have cloned and are cloning the cDNAs for numerous surface antigens
as a first step in determining the structure and function of these cell surface
molecules. Our particular interest is focussed on cell adhesion molecules
expressed on vascular endothelium and by cells of the haemopoietic system in
order to characterise their involvement in the control of the growth and
differentiation of bone marrow stem cells through to mature end cells of the
myeloid lineage.
Julia and Walter Bodmer's Cancer Immunology Laboratory (Julia & Walter Bodmer,
David Bicknell)
The polymerase chain reaction (PCR) has been used in combination with a
human specific Alu primer to develop an assay able to identify individual human
chromosomes in somatic cell hybrids. This application should allow the simple
and rapid characterisation of the human chromosome content in human X rodent
hybrids.
Tony Monaco's laboratory is moving from the ICRF Laboratories at Lincoln's Inn
Fields in London, to the IMM later this year, bringing with him extensive YAC
and genome mapping resources. Chris Higgins' laboratory has an interest in the
formation of the CF and related multi drug resistance genes.
8. University Departments
GENETICS LABORATORY
Head of the Genetics Laboratory and an NHS Consultant in Clinical Genetics.-
Professor John Edwards
My research interests include the theoretical treatment of linkage
analysis and comparative maps of man and mouse. The DNA group (Churchill
Hospital) under Dr A. Miciak is involved in the analysis of a wide range of
clinically significant disorders including Duchenne muscular dystrophy,
Huntington's chorea, neurofibromatosis, retinitis pigmentosa and dystrophia
myotonia.
Garry Brown's Group
Our interests are in clinical genetics and biochemistry of
mitochondrial disorders and the investigation of mitochondrial enzyme
complexes. These overlap and complement those of Dr Gardiner's Group (see
above). The laboratory is an international reference center for studies of
mitochondrial disorders. A major area of research has been on the pyruvate
dehydrogenase complex and characterisation of defects in patients and an
understanding of the complex assembly. We have localised the gene for the E1
alpha subunit to the X-chromosome short arm and have identified the existence
of a second locus on chromosome 4 which encodes a testis-specific form of the
enzyme subunit. We are currently investigating the severity of manifestation
in carrier females which relates both to the nature of the mutation and the
pattern of X-inactivation.
Ian Craig's Group
Our research has concentrated on the molecular organisation of human
sex chromosomes and sex linked disorders, on the carcinoembryonic antigen gene
family and on neurotransmitter enzyme genes.
Current research on the human X and Y is directed to detailed physical
mapping of the regions of surrounding the active steroid sulphatase locus at
Xp23 and the inactive pseudogene on the Y long arm. We are also extending
existing genetic and physical maps for the proximal region of the X short arm
(centered on the hypervariable marker DXS255 at Xp11.22, which we previously
isolated and which has formed a focus for genetic analysis) with particular
reference to the isolation and characterisation of candidate genes for
retinitis pigmentosa, incontinentia pigmenti and Wiskott Aldrich syndrome. A
range of probes is being developed by PCR amplification from irradiated
fragment hybrids and from genomic sequences (YAC and phage clones) isolated
from the monoamine oxidase A and B gene region. Positions of target loci for
disorders are being refined by reference to informative crossovers in families
studied with highly informative markers.
Following on from the isolation and localisation to chromosome 19 of
genomic sequences for the carcinoembryonic antigen (CEA), interest in its
control and the physical and genetic mapping of the super gene families of CEA
and the pregnancy specific glycoproteins have developed under Dr T.C.
Willcocks. Studies on the methylation of coding sequences in the vicinity of
the Myotonic Dystrophy locus in context of imprinting (progressive severity)
are also in progress.
Neurotransmitter enzymes have been studied in collaboration with
Jacques Mallet (Gif sur Yvette) and with John Powell (Institute of Psychiatry,
London). Sally Craig has been involved in mapping a range of such genes
including tyrosine hydroxylase, dopamine beta hydroxylase, tryptophan
hydroxylase and a neurone specific enolase. Collaborative studies on patients
with defects in some of these enzymes are in progress to ascertain their
possible contribution to behavioural disorders.
9. Biochemistry Department
CRC Chromosome Molecular Biology Group (Professor E.M. Southern)
Our aim is to characterise the functional elements of human chromosomes
at the molecular level. We have succeeded in isolating the telomeres of
several human chromosomes as functional ends in yeast (William Brown, Michael
Barnett, Kirsti Kvaloy, Phil McKinnon, Melanie Dobson). The centromere is
likely to be more difficult to isolate, but we have shown that the alpha
satellite maps precisely to the constriction thus providing a molecular
"handle" on the centromere (Chris-Tyler Smith, Katrina Cooper, Neal Mathias).
We are now in a position to design a minichromosome based on these ingredients.
Another area of chromosome molecular biology we are exploring is
targeted integration by homologous recombination using a system that depends on
the interferon induced expression of coding sequences (Andy Porter, Phil Bates,
Jane Ithzaki). We are testing new ways of detecting properly targeted inserts.
We are developing a method for analysing nucleic acid sequences based
on the synthesis of large arrays of oligonucleotides tethered to the surface of
a glass plate. The sequence to be analysed is labelled and hybridised to the
arrays. The pattern of spots which light up, gives information about the
sequence. We have worked out methods for making the array and are now
exploring applications to the analysis of mutations, especially in human
disease (Juliet Honeycombe, Sheila McNutt) and to the analysis of differences
in mRNA populations (Uwe Maskos, Kalim Mir).
We are applying digitizing densitometry and advanced image restoration
procedures to the automatic reading of DNA sequencing gels and to enable
sequences to be read from regions that are too densely packed with bands to be
read by eye (John Elder). Automated reading is also much quicker and less
prone to error than human reading.
10. MRC Immunochemistry Unit (Dr K.B.M. Reid ) Dr R.B. Sim, Dr R.D.
Cambell, and Dr S-K.A. Law.
We are investigating the structure, biological activities, molecular
genetics and gene expression of the complement components and the complement
receptors. Particular interest is focussed on the detailed physical mapping
and sequencing of the human HLA region and the characterisation of amyloid
(amylin) in type II diabetes.
Genetic data suggest that the major histocompatibility complex (MHC)
region spans 3-4 cM. Its size and the existence of a large number of well
characterised loci provide a rationale for the application of detailed physical
mapping which should not only clarify the gene organisation within the complex
region and aid the identification of novel genes, but might also contribute to
understanding the basis for HLA-disease associations. Class I and class II
regions (about 1,500 and 800 kb, respectively) are separated by the class III
region of about 1,100 kb. Large DNA fragment analysis coupled with
identification of HTF islands has enabled the linkage relationships between
major genes of the regions to be established. Deletion studies and physical
mapping have positioned tumour necrosis factors A and B (TNFA & B) between
HLA-B and C2 loci. Population studies have suggested that important disease
susceptibility genes reside within the class III region. Application of the
approach based on the distribution of CpG rich regions has allowed the
identification of a significant number of novel genes including 3 heat shock
protein (HSP70) - related genes. The 390kb separating CYP21B and DRA genes
will be cloned in overlapping cosmids. To date 230kb has already been isolated
and work in progress is completing the characterisation through access to YAC
libraries. Characterisation of cDNAs for the genes mapping to this region will
continue and it is hoped that derived amino acid sequences will yield important
clues as to the function of gene products. Longer term plans include the
determination of the complete nucleotide sequence of the class III region in
collaboration with CEPH (Paris) and the ICRF (London). A major effort will be
invested in the identification of polymorphic markers and use of probes for
"novel" class III genes to examine possible association with various autoimmune
diseases.
Detailed investigations are being carried out on a wide variety of
immune components. Genomic clones for the 3 types of chain (A,B and C) found
in the C1q molecule have been obtained following the initial isolation of cDNA
for the B chain and we have characterised the orientation and order of the
three genes in a 24kb region of chromosome 1. Structure comparisons have
revealed homologies between the C1q chains and several mammalian lectins, which
include mannan binding protein, conglutinin, and lung surfactant protein-A that
may be involved in their similar receptor and complex interactions.
Two C4 genes have been identified (C4A and C4B) and shown to be
separated by only 10kb. They are interesting in that they can differ in size;
C4B is either 22kb or 16kb long depending on the length of intron 9. We have
developed a PFGE technique, that is widely applicable, to identify all
combinations of C4 genotypes so far examined. The complete mRNA sequences for
factor B and C2 have been determined and a number of RFLPs at the gene loci
have allowed further subdivision of the haplotypes carrying certain factor B
and C2 allelic combinations. Deficiency in the cytochrome P450, steroid
21-hydroxylase (CYP21) has been determined to be responsible for congenital
adrenal hyperplasia. Sequencing the presumed defective CYP21B genes in CAP
patients is providing information on nature of the deficiencies. The isolation
of coding sequences for Factor H (the principal soluble non-enzymic regulator
of C3 turnover) has led to confirmation of its localisation close to other
complement controlling genes at 1q31-32. We have also isolated coding
sequences of Factor I (in collaboration with Dr T.J.R. Harris, formerly at
Celltech Ltd.) and have been instrumental in examining deficiencies in this
factor in a number of patients. Long range mapping is in progress and has
extended so far to about 3Mb of the region 4q25-26. Factor I has been
physically linked to the epidermal growth factor gene. Properdin increases the
protective role of complement in conferring resistance to infection. Isolation
of cDNA clones and a genomic clone has allowed sequencing of about 7kb
containing the translated exons. Both mouse and human genes have been mapped,
the latter falling within a region containing the locus for Norrie disease
which is associated with increased susceptibility to infection; however, no
alterations to the properdin locus at Xp11.4 have been observed in such
patients.
The leukocyte adhesion molecules, integrins are a set of heterodimeric
cell surface membrane glycoproteins mediating a range of adhesion activities.
Several subgroups exist and the leukocyte integrins are characterised by a
common beta subunit (CD18 antigen). The gene for CD18 has been isolated and in
situ hybridisation localised the gene to 21q22.1. Sequencing has shown CD18 to
be homologous to a subunit of the fibronectin receptor on chick fibroblasts and
to a range of heterodimeric adhesion molecules whose alpha subunits also share
structural similarities. We are currently studying patients with leukocyte
adhesion deficiency (LAD) caused by defects in CD18; putative point-mutations
will be examined in detail by cloning and sequencing. Determination of the
complete exon/intron boundaries of the CD18 gene has been completed and a
comparison of its control region with related proteins represents the next
major objective.
11. Microbiology
Investigation of cell cycle control in Schizosaccharomyces pombe using
a combination of classical and molecular genetics have identified two major
controls one acting in late G1 leading to the initiation of DNA replication and
the other late in G2 initiating meiosis. The human homologue of one of the
cell cycle control proteins (a protein kinase) has been identified as a
component of MPF. The great significance of the work with S. pombe is the
potential for isolating human homologues of the yeast genes through
complementation studies.
12. Sir William Dunn School of Pathology, University of Oxford, South Parks
Road, Oxford OX1 3RE
Professor Brownlee's Group
In collaboration with the Oxford Haemophilia Centre (Director, Dr C.
Rizza) we have extended the molecular analysis of Factor IX deficiency
(haemophilia B, Christmas disease) to clinical research. Current
investigations are defining the biochemical mechanism whereby a rare group of
patients Hemophilia B Leyden type are unable to express factor IX before
puberty. Studies on the factor IX promotor are in progress and we have
observed that some naturally occurring mutations of the Leyden-type,
haemophilia B patients cause decreased promotor activity in CAT-linked assays
in transformed liver cells. Further studies are examining the detailed
physical map of the region on either side of the factor IX locus at Xq27 and
linking up to defined anonymous loci in the region. This mapping is important
to allow the definition of end-points in deletion patients. Finally, the
disorder will be investigated as a model system for development of therapeutic
approaches.
Peter Cooke and P. H. Dear
We are developing a novel mapping strategy which overcomes the
difficulties of conventional linkage analysis (limited in terms of both
resolution and of informativeness of many markers), using haploid cells and the
polymerase chain reaction (hence "HAPPY" mapping). DNA of a single haploid
cell is broken into large fragments by irradiation, and divided into a number
of aliquots. Each aliquot is then scored for the presence of the sequences to
be mapped, using the polymerase chain reaction. Sequences which lie close
together on a chromosome (relative to the mean size of the fragments) lie on
the same fragment after irradiation, and hence are found in the same aliquot as
each other. Widely separated sequences, conversely, show less tendency to
'cosegregate' in this way. Thus, by analyzing a number of cells the distances
between a pair of sequences may be accurately estimated. By varying the size
of the fragments, the method can be tailored to accurately measure distances
from several megabaseQpairs to a few tens of kilobaseQpairs. We aim to test
this method using three sequences which have already been mapped, then to
extend the method to map many sequences simultaneously.
13. MRC Radiobiology Unit (Dir. Professor G.E. Adams)
This large MRC Unit is situated at Chilton (near Didcot) approximately
15 miles south of Oxford and there are many collaborative links with research
groups in Oxford.
Dr Mary Lyon's Group (Genetics Division)
We are undertaking genetic mapping of phenotypic mutants with
particular reference to those which may be of interest, either as candidates
for effects of developmental genes e.g. homeobox genes, or as potential
homologues of human syndromes. In addition, we are making detailed studies on
genes affecting spermatogenesis and sperm function in the t-complex, in the
proximal region of mouse chromosome 17, including testing candidate genes in
transgenics. A mouse chromosome atlas is maintained, showing the relation of
genetic and G-banded maps, and the positions of segments of homology with human
chromosomes.
Jo Peters' Group/Janet Jones (Genetics Division)
We are interested in the molecular analysis and genetic mapping of
murine biochemical mutants determined by genes with unequivocal homologues in
man e.g. globin, glucose phosphate isomerase, tyrosinase. We are also
analysing by genetic mapping two regions subject to imprinting (distal mouse 2
and proximal mouse 7), and the murine pseudoautosomal region with special
reference to the steroid sulphatase locus. New linkage stocks and PCR
techniques are being developed for rapid mapping of mouse genes and mutations.
The mouse gene list, containing over 2,000 genes is maintained and
published in Mouse Genome, which is compiled and edited in the Genetics
Division. In addition, GeneView, a programme to display genetic maps has been
developed.
Yvonne Boyd's Group (Genetics Division)
Our main interest is in the molecular organisation of the human and
murine X chromosome and in chromosomal inactivation mechanisms. We are using
several approaches to clone novel X-chromosome conserved sequences and have
mapped several genes and many DNA segments onto the human X chromosome.
Conserved sequences and X-linked genes cloned by other groups are being
characterised by detailed mapping in both species using hybrid panels, in situ
hybridisation and an interspecific backcross. We are continuing to develop
hybrid panels and appropriate technologies to simplify their screening (PCR,
non-radioactive in situ hybridisation). Detailed studies of X chromosome
rearrangements and their associated disorders are also in progress e.g.
translocations associated with Duchenne muscular dystrophy in females (with
Genetics Laboratory) and a microdeletion of the mouse X chromosome, Ta25H.
Dr A. G. Searle (Genetics Division)
I maintain and update a databank (in dBase III) of homologous human and
mouse genes, including anonymous DNA segments, which have been assigned to
chromosomes in both species. Over 500 such homologies are now listed and
printouts are available in a variety of forms.
Dr John Thacker (Cell and Molecular Biology)
We are mapping large deletions which include the hypoxanthine
phosphoribosyl transferase locus in human cells with the use of pulsed field
gel electrophoresis and and a variety of probes from the Xq26 region. We are
also establishing methods to map and to isolate DNA repair genes by
complementation of radiosensitive mutants.
5. A Strategy for Genome Analysis
Hans Lehrach
Imperial Cancer Research Fund
PO Box 123
Lincoln's Inn Fields
London WC2A 3PX
The genome of an organism contains all the information necessary to
establish its features and capabilities. Direct analysis of this information
might therefore offer a way to approach many biological and medical problems
over a new route.
The ideal data set for this work consists of the DNA sequence of the
genome (3.109 basis for the human), information on sequence conservation, as
well as information on the pattern of expression of this sequence information
in different tissues and stages of development. Different approaches have been
proposed to determine a part of this information, ranging from the
determination of the entire sequence of the human genome, over proposals
relying more heavily on mapping (possibly based on the use of sequence tagged
sites) to an emphasis on the sequencing of transcripts (the British genome
project).
Our work has started to develop an alternative approach, which we
believe will allow an efficient determination of much of the required
information. This proposal relies heavily on the use of very informative
hybridisation probes (e.g. short oligonucleotides) to many clones in parallel,
made possible by the use of a robotic device to spot more than 9000 clones
stored in microtiterplates in a high density array on nylon membranes. In
addition the use of these high density filters opens the way to use these
libraries as a high density molecular mapping system, and therefore allows the
easy establishment of data bases containing information collected by different
techniques in different laboratories.
We expect this approach to develop in four stages:
1) The development of large insert (cosmid, P1 and YAC) libraries into a global
mapping system of molecular resolution (reference libraries), based
predominantly on providing high density replica filters of the stored libraries
and accumulating the information on clones and probes in a relational data base
format.
2) The transition from the maps in arbitrary coordinates (coordinate of clone)
or coordinates within a contig or ultimately a chromosome based on the ordering
of clones into long contigs by hybridisation-fingerprinting, using "relatinal"
mapping of clones in multiple libraries (coxmid, P1, YAC, radiation hybrids
etc.).
3) Sequence-fingerprinting of cDNA clones and of short (app. 2 kb) genomic
clones of homologous regions (the X chromosome?) in two closely related
organisms (man and gibbon), using 6 to 8 mers as hybridisation probes, using
conditions allowing the identification of perfectly matched hybrids. (Drmanac
et al, in press). Information on similarities and differences in hybridisation
patterns should identify identical or similar cDNA clones, locate cDNAs on the
genomic sequence and identify the conserved sequences in the corresponding
chromosomes.
4) And finally complete (or close to complete) sequence analysis, minimally of
the conserved sequences (mostly exons) identified at the previous stage using
standard sequencing techniques, or alternatively the determination of
essentially complete sequences of chromosomes or genomes by exhaustive
hybridisation sequencing (Drmanac et al, 1989).
As part of the first stage, the development of systems to provide easy
access to libraries and their development into a molecular mapping system, we
have constructed a number of chromosome specific cosmid libraries from flow
sorted chromosomes provided by Brian Young (ICRF). Libraries from human
chromosomes X and 21 are available at the moment, while additional libraries
are under construction (chromosomes 17, 22 and 11 will be available in the near
future) (Nizetic et al, unpublished). Libraries are picked into microtiter
plates, spotted in high density grids (currently app. 9000 clones per filter)
and converted into DNA spots. In addition to being used in hybridisation-
fingerprinting experiments designed to establish ordered clone libraries, these
filters have been provided to over 30 laboratories in many countries, and have
been used there to identify clones hybridising to more than 100 different
probes from these chromosomes. Hybridisation results have been sent back to us
together with some information on the probe used, positive clones are
identified, sent back to the originating laboratory, and data on clone and
probe are entered into a relational data base (currently based on Oracle).
In addition, we hope to establish a similar (global) clone
identification and mapping system based on human (Monaco, unpublished) and
mouse YAC libraries (Larin, unpublished), since the large average insert size
of the libraries (650 to 700 kb) should allow us to construct high density
filters with minimally two genomes per filter.
The second stage, the ordering of the libraries by hybridisation
fingerprinting, relies mainly on the use of short oligonucleotides as
hybridisation probes (Nizetic et al., unpublished), complemented (for the large
insert libraries and at the later stages of the analysis) by the use of pools
of unique probes (cDNA clones, probes from the ends of cosmid clusters or
YACs). The approach has been tested by theoretical analysis (Michiels et al.,
1987, Lehrach et al., in press) and computer simulation (Poustka et al., 1986).
In addition an experimental test has been described (Craig et al., 1990). To
test the interplay of the different experiments with the analysis and data base
programs, we are currently analysing the genome of S. Pombe, a biologically
interesting organism with a small genome and few repeated sequences. In
addition to this first large scale test of the hybridisation fingerprinting
protocol on an unknown genome we have started work on the analysis of libraries
covering the human X chromosome, chromosome 21 and the genome of D.
Melanogaster (Hoheisel et al., unpublished).
Similarly we expect to be able to use hybridisation fingerprinting to
order YAC libraries covering the human and mouse genome. In this case we will
also try to use pools of cDNA clones as hybridisation probes, since this should
allow a concurrent mapping of both genomes in parallel, and might permit the
combined use of techniques from mouse genetics and human somatic cell genetics
to relate the maps to each other and to the genetic maps of both organisms.
The third stage of the analysis, the use of
hybridisation-fingerprinting of short clones from cDNA libraries and genomic
libraries has been tested successfully in the analysis of three hundred cDNA
clones (Drmanac et al, unpublished). DNA from the clone inserts was amplified
by PCR with flanking primers, spotted on filters, and analysed by hybridisation
with hepta or octa oligonucleotides under conditions discriminating against
imperfect matches. Larger scale tests involving the analysis of large human
and mouse cDNA libraries are planned. Due to the high efficiency of the
sequence-fingerprinting protocol even large projects (e.g. the analysis of a
human chromosome) should be well within the theoretical capacity of fairly
small groups.
In contrast to the first three stages described above, we expect the
fourth stage of the analysis, essentially the determination of the complete
sequence of a human chromosome, to require either considerable further progress
in the technique (higher density of DNA spots on the filter, further automation
of the hybridisation and reading steps), or a considerably larger scale (and
cost) than the first three stages.
Though difficulties and unexpected bottlenecks might appear at
different steps of the analysis, we expect that the use of hybridisation
techniques as fingerprinting and ultimately sequencing protocol, in combination
with the high efficiency of the use of common libraries as mapping resources,
will increase speed and cost effectiveness of analysing genomic and cDNA
information significantly, and might be able to achieve the ten fold or higher
increase in speed postulated in some of the scenarios of analysing the human
genome.
Acknowledgements
I would like to thank Rade Drmanac, Jurg Hoheisel, Dean Nizetic, Tony
Monaco, Zoia Larin, Greg Lennon and Gunther Zehetner as well as the technical
assistants involved in this work for their contribution to the development of
the concepts, and their execution. In addition I would like to thank Annemarie
Poustka and members of her laboratory for discussions and experimental results,
and my colleagues at the ICRF for discussions.
References
Craig, A. G., Nizetic, D., Hoheisel, J. D., Zehetner, G. and Lehrach H.
(1990) Ordering of cosmid clones covering the Herpes simplex virus type 1
(HSV-1) genome: a test case for fingerprinting by hybridisation. Nucl. Acids
Res. 18, No 9; 2653
Drmanac, R., Lennon, G., Drmanac, S., Labat, I., Crkvenjakov, R. and Lehrach,
H. (1990) Partial sequencing by oligo-hybridization: concept and applications
in genome analysis. In press
Lehrach, H., Drmanac, R., Hoheisel, J., Larin, Z., Lennon, G., Nizetic, D.,
Monaco, A., Zehetner, G. and Poustka, A. (1990) Hybridisation fingerprinting
in genome mapping and sequencing. In press.
Michiels, F., Craig, A., Zehetner, G., Smith, G. P. and Lehrach, H. (1987)
Molecular approaches to genome analysis: a strategy for the construction of
ordered overlapping clone libraries. CABIOS 3; 203
Poustka, A., Pohl, T., Barlow, D. P., Zehetner, G., Craig, A., Michiels, F.,
Ehrich, E., Frischauf, A.M. and Lehrach, H. (1986) Molecular approaches to
mammalian genetics. Cold Spring Harbor Symposia on Quantitative Biology,
Volume LI; 131.
6. Directed Programme awarded project grants (as at 1.2.90)
Furzana Bayri
Medical Research Council
20 Park Crescent
London W1N 4AL
Application Project
Dr. N.A. Affara, Dr. M.A. Yuille Towards a 1-centimorgan map of
and Prof. M.A. Ferguson-Smith chromosome 9 using dinucleotide
(Pathology, Cambridge) polymorphisms and meiotic
recombination in sperm
Dr. Donna Albertson Non-isotopic in situ hybridization
(MRC Molecular Genetics Unit, mapping of human genomic and
Cambridge) cDNA clones to chromosomes and
tissues
Dr. J. Bell, Dr. K. Davies PCR analysed polymorphisms for
and Dr. J. Todd the construction of a genetic
(Nuffield Dept. of Surgery, linkage map of the human genome
John Radcliffe Hospital,
Oxford)
Dr. D.R. Bentley, Prof. F.B. Giannelli Long-range mapping of regions of
and Prof. M. Bobrow the X-chromosome using YAC and
(Paediatric Research Unit, cosmid overlapping
United Medical & Dental Schools
of Guy's and St. Thomas's Hospitals,
London)
Dr. H. Beswick, Dr. A.M. Goate and Isolation and sequencing of human
Prof. R. Williamson disease loci
(Biochemistry and Molecular
Genetics, St. Mary's Hospital
Medical School, London)
Dr. E. Boyd & Prof. J.M. Connor Chromosomal mapping using in
(Duncan Guthrie Institute of situ hybridisation with
Medical Genetics, Glasgow) biotinylated DNA sequences
Dr. S.D.M. Brown and The molecular mapping of the
Dr. K. Johnson myotonic dystrophy (DM) locus
(Biochemistry and Molecular in mouse and man
Genetics, St. Mary's Hospital
Medical School, London)
Dr. V.J. Buckle & Prof. Sir David The use of confocal laser
Weatherall (Institute of scanning microscopy in human
Molecular Medicine, Oxford) genome mapping
Prof. M. Bobrow, Prof. F. Walsh The use of oligonucleotides in
and Dr. D. Bentley studies of genome mapping and
(Medical & Molecular Genetics, for analysis of polymorphisms
Guy's Hospital, London) and mutations in human genetic
diseases
Dr. B.M. Cattanach, Dr. Y.L. Boyd Genetic and physical mapping
and Dr. M.F. Lyon studies
(MRC Radiobiology Unit, Oxon)
Dr. J.F. Collins and Dr. A.F.W. Computational tools for the
Coulson Human Genome Project
(Biocomputing Research Unit,
Edinburgh)
Dr. H. Cooke A comparative and functional
(MRC Human Genetics Unit, analysis of mouse telomeres
Edinburgh)
Dr. J.D.A. Delhanty & Dr. M.S. Povey Precise mapping of human
(Galton Laboratory, Genetics & sequences using fluorescent
Biometry, UCL, plus MRC Human in situ hybridisation
Biochemical Genetics Unit, London)
Dr. J.K. Elder and Prof. E.M. Southern Image processing in genome
(Biochemistry, Oxford) analysis
Prof. H.J. Evans, Dr. N.D. Hastie, A DNA extractor and gel
Dr. C.M. Steel & Dr. H.J. Cooke documentation system for
(MRC Human Genetics Unit, human gene mapping
Edinburgh)
Prof. H.J. Evans, Dr. A.F. Wright Mapping of human genetic
and Dr. M.G. Dunlop disorders using short tandem
(MRC Human Genetics Unit, repetitive (STR) loci by
Edinburgh) multiplex amplification and
oligonucleotide probing
Prof. M.A. Ferguson-Smith, Human gene mapping and
Dr. N.A. Affara & Dr. N.P. Carter sequencing
(Pathology, Cambridge)
Prof. P.S. Harper and Dr. D.J. Shaw Gene mapping of inherited
(Institute of Medical Genetics, neurological disorders
Cardiff)
Prof. H. Harris, Prof. G. Brownlee Equipment for mapping the
and Dr. P.R. Cook human genome
(Sir William Dunn School of
Pathology, Oxford)
Dr. P.C. Harris and Dr. D.R. Higgs The construction of a physical
(Institute of Molecular map of the human chromosome
Medicine, Oxford) band 16p13.3
Dr. N.D. Hastie, Dr. I.J. Jackson Developing procedures for
and Dr. V. van Heyningen introducing YACs with
(MRC Human Genetics Unit, mammalian inserts into
Edinburgh) cultured cells and mice
Dr. N.D. Hastie, Dr. V. van The construction of mouse
Heyningen, Dr. I.J. Jackson models of human disease and
and Dr. D.J. Porteous the use of CpG island traps
(MRC Human Genetics Unit,
Edinburgh)
Dr. R. Hill, Dr. N.D. Hastie Large genomic fragment
and Dr. D.R. Davidson cloning in YAC libraries for
(MRC Human Genetics Unit, long range genetic analysis
Edinburgh) and isolation of mouse mutations
Dr. D.A. Hopkinson Evaluation of denaturing
(MRC Human Biochemical gradient gel electrophoresis as a
Genetics Unit, London) routine procedure for use in
human gene mapping
Dr. D.A. Hopkinson Equipment for multiple DNA
(MRC Human Biochemical processing and analysis in human
Genetics Unit, London) gene mapping
Dr. D.A. Hopkinson Analysis of data relating to
(MRC Human Biochemical human gene mapping
Genetics Unit, London)
Dr. K. Kaiser and Application of a new subtraction
Prof. J.M. Connor hybridisation procedure to the
(Genetics, Glasgow) cloning of the gene for human
congenital adrenal hypoplasia
Dr. D.R.F. Leach Improved hosts for cloning
(Molecular Biology, human DNA sequences
Edinburgh)
Dr. P.F.R. Little Cosmid fingerprinting mapping of
(Biochemistry, Imperial College, the short arm of human
London) chromosome 11
Dr. P.F.R. Little, Dr. J. Sulston Random cloning vectors
and Dr. A. Coulson
(Biochemistry, Imperial College,
London and LMB, Cambridge)
Prof. G. Lowe and Dr. P.R. Cook Development of reagents that
(Dyson Perrins Laboratory, cleave DNA at any predetermined
Oxford) site
Dr. S. Malcolm Long range mapping of
(Mothercare Department of chromosome 15 and the X
Paediatric Genetics. Institute chromosome
of Child Health, London)
Dr. G. Melmer and Dr. H. Gurling Gene detection by oligonucleotide
(Psychiatry, University College hybridisation
and Middlesex Medical School,
London)
Dr. J. Peters and Dr. Y.L. Boyd Improved resources for genetic
(MRC Radiobiology Unit, and physical mapping in the mouse
Didcot, Oxon)
Dr. B. Ponder, Mapping the pericentromeric
(Pathology, Cambridge) region of chromosome 10
Dr. D.J. Porteous, Dr. V. van High resolution mapping of
Heyningen and Dr. J.R. Gosden human chromosomes
(MRC Human Genetics Unit,
Edinburgh)
Dr. T.H. Rabbitts and Dr. G.P. Winter Sequence analysis of the human
(MRC, LMB, Cambridge) immunoglobulin heavy chain
variable region gene locus
Dr. N.J. Royle The organisation of mini-
(Genetics, Leicester) satellite sequences in the
proterminal region of the long
arm of chromosome 7
Prof. E.M. Southern and Mr. J.K. Elder Application of phosphor
(Biochemistry, Oxford) imaging in genome analysis
Dr. C. Tyler-Smith and Analysis of human centromeres
Dr. W. Brown and telomeres
(Biochemistry, Oxford)
Dr. P. Whittaker and Dr. I. Day Improved strategies for genomic
(Clinical Biochemistry, walking and cDNA library sorting
Southampton)
Dr. J. Wolfe and Dr. S. Povey A general method to isolate
(Genetics and Biometry, UCL) probes and genes from small
regions of the human genome:
its application to mapping 9q
Dr. A.F. Wright Construction of large-insert
(MRC Human Genetics Unit, human yeast artificial
Edinburgh) chromosome libraries
Prof. E.M. Southern Development of DNA analysis
(Biochemistry, Oxford) methods
Directed Programme Grants
For your information the dates for the next Directed Programme
Committees and the subsequent deadlines for any project grant applications are
as follows:
Directed Programme Deadline for Project Grant
Committee Applications
1st November 1990 3rd October 1990
27th February 1991 29th January 1991
21st May 1991 22nd April 1991
7. Provision of expenses for attendance at HGMP meetings
The following details the guidelines and conditions of the scheme
whereby applicants may have expenses, relating to HGMP meetings, reimbursed by
the Medical Research Council.
1. HGMP funds for attendance at meetings will be restricted to:
1.1 Recipients of grants and those supported on grants awarded through
the HGMP Directed Programme.
1.2 Holders of HGMP Fellowships and Studentships.
1.3 Recipients of direct support through the HGMP.
1.4 Invited participants/chairpersons at HG business meetings (eg.
chromosome workshops, etc.)
2. Those fulfilling the eligibility requirements set out in 1 may apply for:
2.1 Full costs for UK participants at relevant meetings held in the UK
(travel, accommodation, subsistence, etc.).
2.2 Accommodation/registration costs for non-UK participants at
relevant meetings held in the UK.
2.3 Travel costs, and other essential expenses not covered by the
organizers, for UK participants at relevant meetings held overseas.
2.4 Expenses for attendance of UK participants at meetings arranged by,
or formally sponsored through, the UK HGMP initiative.
3. Factors to be taken into consideration when reviewing applications will be:
3.1 that the meeting is directly relevant to genome mapping;
3.2 that each centre or laboratory is adequately rather than
excessively represented at any given meeting;
3.3 that the UK is properly represented at international meetings.
4. Additional conditions applicable to requests for funds will be:
4.1 that individuals be encouraged to apply for all or part of the
expenses from their parent Institution, or elsewhere, should this be
appropriate;
4.2 that in normal circumstances, any one individual may only apply for
funds to attend one national and one international meeting per year;
4.3 that young investigators be given preference for funding, on the
premise that established workers can usually secure support from elsewhere.
5. Applications for funds to organise relevant HGMP meetings in the UK should
be submitted as fully costed proposals for consideration by the Directed
Programme Committee.
8. User Registration for access to the services of the Resource Centre
Mrs. Christine Bates (Administrator)
HGMP Resource Centre
Clinical Research Centre
Watford Road
HARROW, Middx, HA1 3UJ
Any UK "academic" research worker with interests relevant to the
objectives of the UK Human Genome Mapping Project can apply for registration as
an HGMP user of the Biological and/or Computing Services. It is not necessary
to have support from the MRC or the HGMP Directed Programme.
Foreign or non-academic users, or those whose field of work is not
judged to be relevant to the interest of the HGMP can also apply for
registration, but their access may be subject to charging or restrictions.
The forms for registration can be obtained from the Resource Centre
(081 869/3808 or 3446). Many units, institutions and individuals here already
have been sent the form together with details of some of the services. If a
number of people in any of the various organisations wish to register please
copy the form.
In the first instance the aim is to provide the following services:-
a) access on-line to e-mail and bulletin board facilities, and to a wide range
of standard genetics and molecular biology software packages.
b) computer training courses at the Resource Centre in 1991, although
pilot-courses have been run in Cambridge during this year.
c) access to the Human Cell Bank set up at CAMR at Porton the current service
comprising:-
(i) EBV-transformation.
(ii) distribution of human cell-lines from the collection.
(iii) testing for and eradication of mycoplasma infection.
(iv) a depository for irreplaceable cell stocks.
(v) supply of cell-pellets for DNA extraction and/or preparation and
supply of the DNA.
d) supply of DNA probes as listed in the UK DNA Probe Bank catalogue. This
distributive service has recently transferred from ICRF Clare Hall Laboratory.
Other services planned for the near future include the synthesis of
oligonucleotides and a YAC screening resource.
To date many researchers have already registered and we would urge
other interested parties to do so as soon as possible. In particular, could
current users of extant services, such as the Human Cell Bank or the UK DNA
Probe Bank, please register by the deadline of 1st November 1990.