Effect of rare codons on gene expression

Laura K. Moen, Ph.D. LKM100F at ODUVM.CC.ODU.EDU
Mon Jun 26 14:55:36 EST 1995

Hi, guys - I am reposting because our usenet did not "get" any of lastst
week's posts, so I may have missed some of your replies.  Please repost
your replies if possible or EMail me this time.  Thank you.  LKM
----------------------------Original message----------------------------
Hi, Netters.
  I have been attempting to clone and express a gene from a eukaryotic
virus in E. coli.  I appear to have been successful, but I am not
getting the same levels _consistently_ and overall, expression is not
as high as I had hoped it would be.  Aside from numerous other factors
which I am addressing, I am curious as to what folks out in the net
think about the effect of codon use on gene expression.  I have heard
various anecdotal stories about the genes cloned with rare codons and
high expression levels (or at least reasonable ones) were still obtained.
The book by Gold and Reznikoff on Gene Expression, however, has some
compelling evidence to suggest otherwise.  At present, my expression
levels are so low as to make detection with Westerns OK, but not very
abundant (alkaline p'tase detection).  Is it worthwhile to consider the
expense and time of synthesizing the gene with more abundant codons?
Thanks for any input.  Post or Email to LKM100F at ODUVM.cc.odu.edu

Laura K. Moen, Ph.D.
Associate Professor
Dept. Chemistry & Biochemistry
Old Dominion University
Norfolk, VA 23529-0126


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