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express protein with many rare codons


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#1 saad

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Posted 01 February 2010 - 11:06 PM

Hi all

I'm trying to express domain/fragment of protein ligated to pGEX-6P.
this sequence contains many rare codone for example:
CGG GGG = 1
CGG CGG CGG = 1
CGG CCC = 1
they are about 15 codons. I tried in BL21 (DE3) and Rosetta but it seems there is nothing.
I tried different temp and IPTG conc. but unfortunately I can't. when I checked the insoluble fraction I found a band at the right size of my protein but I don't know from the whole story if it easy to express protin like this one.
any suggestions or guide I will appreciate any reply.

Thanks in advance

#2 scolix

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Posted 27 February 2010 - 01:17 PM

Are these codon optimised for bacterial expression? Sometimes it might be better to check if the cDNA has been optimised for a specific expression system.
This might not be the reason for no expression but could help.




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