SOFER@BIONET-20.ARPA (02/05/88)
From: William H. Sofer <SOFER@BIONET-20.ARPA>
Rapid Cloning of Genes
Encoding Sequence-specific DNA Binding Proteins
A Workshop in Molecular Genetics and Biotechnology
Presented by
Drs. Steven L. McKnight, Karen LaMarco and Charles Vinson
April 21-22, 1988
At the Waksman Institute, Rutgers University
Piscataway, NJ 08854-0759 201-932-4258
This course is intended for those actively engaged in the
field of eukaryotic gene regulation, and who are interested
in a new (unpublished) technique for the cloning of genes
that code for proteins that bind to DNA.
One of the goals of those studying gene regulation is to
identify and purify proteins that bind to DNA in a sequence-
specific manner. These trans-regulatory proteins, by binding
to cis-regulatory DNA sequences located within promoter and
enhancer elements, play an important role in the regulation
of gene expression.
Dr. McKnight and his colleagues have developed a technique
that has allowed them to screen directly for lambda gt-11
clones that encode proteins that bind to specific DNA
sequences. Their procedure, which involves several
simple modifications of standard lambda gt-11 screening,
offers a potentially rapid method for the isolation of genes
that encode rare DNA binding proteins.
The two day course will cover a start-to-finish screening
procedure, offering hands-on experience with DNA probes,
filter lifts and treatment of culture plates. At the end of
the course each participant will be provided with a sample of
the lambda gt-11 bacteriophage that encodes the protein that
serves as a positive control.
-------otto@CSHL.ORG (Karen Otto in the Meetings Office) (12/08/90)
COLD SPRING HARBOR LABORATORY
SPRING COURSE ANNOUNCEMENT
PROTEIN PURIFICATION AND CHARACTERIZATION
April 8 - 22, 1991
Daniel R. Marshak, Cold Spring Harbor Laboratory
Bruce W. Erickson, University of North Carolina
Jim Kadonaga, University of California, San Diego
John A. Smith, Harvard Medical School
This course is intended for scientists who are not familiar with
techniques of protein isolation and characterization. Students will
learn the major techniques in protein purification by actually
performing four separate isolations including: (i) a regulatory
protein from muscle tissue; (ii) a fusion protein from E. coli;
(iii) a DNA binding protein from nuclei of tissue culture cells; and
(iv) a chemically synthesized peptide. A variety of
chromatographic, electrophoretic, and bulk fractionation techniques
will be employed including: ion exchange, gel filtration,
hydrophobic interaction, affinity-based adsorption, and
immunoaffinity chromatography; polyacrylamide gel and two-
dimensional gel electrophoresis and electroblotting; precipitation
by salt and pH; and HPLC analysis. Methods of protein
characterization will be discussed including amino acid analysis,
protein sequencing, and mass spectrometry. Emphasis will be placed
on methods of protein purification rather than automated
instrumental analysis. Guest lecturers will discuss protein
structure, modifications of proteins, and methodologies for protein
purification. Applications of protein biochemistry to various areas
of research in molecular biology will be discussed. Guest lecturers
include: R. Aebersold, L. Gierasch, G. Hart, Y. Paterson, N. Pace,
G. Rose, and K. Wilson.
* * * * * * * * * * * * * * * * * * * * *
CLONING & ANALYSIS OF LARGE DNA MOLECULES
April 8 - 22, 1991
Bruce Birren, California Institute of Technology
Sue Klapholz, Cell Genesys, Inc.
Nancy Shepherd, E. I. du Pont de Nemours & Co.
This course will cover the theory and practice of manipulating and
cloning high molecular weight DNA. Lectures and laboratory work
will deal with the use of bacteriophage P1 and yeast artificial
chromosome (YAC) cloning systems, the isolation and manipulation of
high molecular weight DNA from mammalian cells for cloning
(including the size-selection of >200 kb DNA fragments), and the
analysis of high molecular weight DNA by pulse field gel (PFG)
separation techniques. P1 and YAC recombinant DNA molecules will be
produced, introduced into cells (E. coli and yeast, respectively),
and reisolated after appropriate clone selection and colony
screening procedures. A variety of size standards for pulsed-field
gel electrophoresis will be prepared and gels will be run to compare
the DNA separation capabilities of the common PFG techniques.
Students will gain experience with physical mapping of YAC inserts
and high molecular weight genomic DNA. Lectures by outside speakers
on topics of current interest will supplement the laboratory work.
Application deadline: January 15, 1991
Tuition, Room and Board . . . . . . . . . . . . . . . . . . $1,475
Application/information may be obtained from:
REGISTRAR
Cold Spring Harbor Laboratory
Bungtown Road
Cold Spring Harbor, NY 11724
EMAIL: needinfo@cshl.org
(include your name & address as well as course name)
PHONE: (516) 367-8346
FAX: (516) 367-8845kristoff@GENBANK.BIO.NET (David Kristofferson) (04/26/91)
SHORT COURSE IN BASIC TISSUE CULTURE
presented by
The Biotechnology Program
Manhattan College/College of Mt. St. Vincent
Bronx (Riverdale), NY
JUNE 3 - 6, 1991
The essential aspects and contemporary methods of growing
cells in culture will be presented mainly by hands-on procedures
and some demonstrations. There will be lectures on the cultured
cell, monolayer and suspension culture systems, cryogenic preser-
vation, primary cell culturing, contemporary methods of cell
identification, and cell nutrition. Hands-on procedures will
include pipetting, media preparation and filtration, the hemocy-
tometer method of cell counting, cryogenic preservation, cultur-
ing anchorage dependent cells, primary cell culturing, cell
identification using immunohistofluoresence, and karyotyping.
Demonstrations on the use of cryogenic equipment, various types
of culture vessels, two dimensional gel electrophoresis of cul-
tured cell proteins emphasizing labeling, isoelectric focusing,
polyacrylamide gel electrophoresis and autoradiography will also
be presented.
GUEST FACULTY
John Ryan
University of Connecticut
Dr. Peter Del Vecchio
Albany Medical College
Dr. James Haley
College of Mt. St. Vincent/
Einstein College of Medicine
For additional information contact the Course Director:
Frances M. Cardillo, Department of Biology, Manhattan College
Mount St. Vincent Campus, Bronx, New York 10471
PHONE: 212-549-8000 ext367 FAX: 212-549-0915
Bitnet: ARF6@MANVAX.BITNET