Looking for dnaY gene

Guy Oshiro Guy.Oshiro at UCHSC.edu
Thu Sep 19 10:33:13 EST 1996


Dear Netters,

   I am currently trying to express my eukaryotic gene (my favorite
gene - MFG) in a pGEX system in E. coli.  I am having great difficulty
to get adequate expression of this fusion protein following induction. 
I am making enough fusion protein to see it on a western but not enough
to see it by Coomasie stain.

Analysis of MFG showed many rare Arginine codons AGA and AGG and
several tandem repeats. Upon doing a literature search - I discovered
several references that mentioned that having tandem repeats of AGA and
AGG codons can cause premature termination.  In fact this is what I see
on my western blots - premature termination of my fusion protein with
distinct bands that correspond to the location of the tandem repeats.

Anyways, let me get to the point of this posting.  These researchers
showed that they could get high expression of their proteins with
tandem repeats of AGA and AGG codons by supplementing with the dnaY
gene that codes for the Arg tRNA in E. coli.  I am looking for a source
for this dnaY gene, plasmid pUBS520 in the article.  Or any other
derivative.

Thanks in advance for the help.

Guy Oshiro
EMAIL: Guy.Oshiro at UCHSC.edu

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