May I just use conventional E.Coli strain? - (Nov/29/2005 )

I am going to clone a cDNA whose product is a transmembrane protein and potentially harmful to bacteria. Someone suggest me use some recombination-deficient E.coli strain, such as SURE. But if my cDNA is in a vector which drive expression only in mamalian system, may I still use regular E.coli strain to do my work?

Is the gene mammalian? Does the protein have to be glycosylated etc? If its a mammalian genes its unlikely to be expressed and then the translated protein folded correctly in a bacterial system. If the promoter is only recongised by mammalian system then I think you can be safe that it wont be expressed in E. coli. I think the short answer is, there is no way of knowing unless you try it.

hi
ML1975 is not wrong. You will know the answer by trying it.
But assuming promoter controlling the cDNA is a mammalian one, you won't encounter particular problems with your amplification.