EROSENTHAL@BIONET-20.BIO.NET (Eric Rosenthal) (01/14/89)
My lab, and other labs at Kewalo Marine Lab, have been trying
to get expression of cloned genes using the Bluescript vector
(plasmids SK- and SK+). We have constructed a variety of
recombinants using different restriction fragments in different
restriction sites. All of these should have the cloned DNA in the
correct reading frame, but we have not been able to detect the
synthesis of any new proteins following IPTG induction. We have
tried various strains of e. coli--JM101, DH1 and XL1-blue. Strategene,
which makes the Bluescript vectors, has suggested that we should
continue to try different strains of bacteria until we find one that
works. We would be interested to hear from other people who have
experience with Bluescript or other expression vectors. Is it
really necessary to randomly fish around using different strains of
e. coli until you find one that works? Is there a special trick to
getting proper induction? Any suggestions will be greatly
appreciated.
Eric Rosenthal
EROSENTHAL@BIONET-20.BIO.NET
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