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Cloning two genes in the same plasmid with one promotor - (Apr/08/2014 )

Hello!

I want to coexpress two different genes in E. coli.  Is it possible to clone and coexpress two genes, that are part of the same operon and overlap, in the same MCS of pET26b ?

The idea is to get a polycistronic mRNA and two different proteins: One with a His-Tag and another one without. 

 

-Flo-

-Flo1988-

This should work.

-phage434-

Just curious, would you need to make sure to place a shine-dalgarno/rbs before each gene?

-skepticalguy90-

That would be a good idea. It certainly would improve expression of the second gene.

-phage434-

I think the first gene already contains a SD Sequence for the translation of the second gene.

Thanks for your replies

-Flo1988-