Cloning vector for high GC ratio gene? - is GC ratio a criteria in choosing vector??

17 May 2016 1 5K Report

I'm trying to express a my target gene which has a high GC ratio about 75% in E. coli. The thing is, I'm not sure if the ratio of GC is a criteria in choosing the vector or rather, if there are specific plasmids that are more suited to expression of these sort of genes. Any suggestions?

Thanks!

Jan Claesen

Dear Hossam,

Your cloning efficiency should not be affected by the %GC content of the insert you wish to introduce. However, PCR amplification can be more difficult (try adding DMSO to your reaction if you are having trouble) and the gene might not express (efficiently) in E. coli because of inefficient translation (which can be solved by codon-optimizing your gene of interest to match E. coli's preferences).

Best regards,

Jan

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