Take the 2-minute tour ×
Biology Stack Exchange is a question and answer site for biology researchers, academics, and students. It's 100% free, no registration required.

When I express proteins in bacteria I put at least two stop codons at the end of the gene to increase the termination efficiency. Is this the case in eukaryotic cells too? If I put a single stop codon is there a risk for the ribosomal complex to readthrough and continue translation?

share|improve this question
    
There is no such practice as far as I know. Personally I have never done that and things work fine. You can add a polyadenylation signal, though. Is this a synthetic gene or something ? –  WYSIWYG Apr 10 '13 at 4:06
1  
From what I understand the eRFs are not perfect but they still work better than RF1 and RF2 which allow for a non trivial amount of readthrough. –  bobthejoe Apr 10 '13 at 5:09
    
This is a synthetic gene. I already designed a primer with 3 stop codons at the end of the gene. I will post here again if I see any improvement in gene expression (with compared to other similar constructs in our lab). –  Engin Yapici Apr 10 '13 at 19:28

1 Answer 1

There is a paper for transfection of mammalian cells here which has a bit of comparison of stop codon and protein yield.

Otherwise this article suggests having a 'rare' codon after the stop to prevent readthrough. But yes, I'm not sure why one wouldn't simply put two stops after each other.

share|improve this answer

Your Answer

 
discard

By posting your answer, you agree to the privacy policy and terms of service.

Not the answer you're looking for? Browse other questions tagged or ask your own question.