response: cDNA expression library screening

Dan Vernon vernondm at whitman.edu
Fri Oct 10 17:30:29 EST 1997


Zhixiang Chen:   Regarding your question about unexpected expression of
plant mRNAs in your expression library--> Lots of plant genes could have
fortuitous E. coli ribosome binding sites upstream of an ATG:  the
consensus ribo binding site is something like AAGGAG-  but as little as
AGGA or GAGG will allow some translation.  I once was able to massively
express a plant protein in E. coli by simply plopping the full-length
cDNA into bluescript behind the T7 promoter, then transforming an E.
coli strain that expressed T7 RNA pol.ase. The cDNA had an AAGGAG-like
sequence in the 5'UTR just upstream of the initiation AUG.
   So, what you suggest is happening in your library is very possible.
-Dan Vernon

Department of Biology
Whitman College
Walla Walla, WA 99362
vernondm at whitman.edu
509-527-5326






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