Internal ribosome binding sites.
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.
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
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