Internal ribosome binding sites.

Warren Wakarchuk warren at nrcbs8.bio.nrc.ca
Wed Sep 7 10:03:11 EST 1994


  I have been working with a gene which gives me minimal expression in an 
expression vector that has been used for numerous other genes to give 
excellent production.  I ran across an old posting describing the effect of 
tandem rare (AGA, AGG)  arg codons and how this gives rise to false 
ribosome binding sites.  On inspection of my sequence I did not find the arg 
codons but instead a sequence which looks like a pretty good RBS to me.

SEQUENCE  AAAGGAGAA

My query is this:  Is is worth while to go in and mutate the sequence to see 
if the expression levels go up, or is this idea of internal RBS sites messing 
up translation a long shot.

FYI : the gene is 290 aa, very AT rich (60%), and does not appear to be toxic, 
does not make inclusion bodies, it can only be detected with antibodys ie no 
big band on SDS-PAGE.  The vector is a pTAC/F1ori/lacI/bla containing pBR322 
derivitive.

Any thoughts? 


.



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