Protein expression with T7 promoter (View forum version)



Spright

Posted 29 December 2009 - 05:19 PM

Hi all,

Many of us use the E. coli BL21 (DE3) strain to express cloned genes that have a T7 promoter.

DE3 strains are lysogenic for a λ prophage that contains an IPTG-inducible T7 RNA polymerase.

My question is: what if I would like to express my protein in bacteria other than E. coli? (Shigella in my case). Are there similar commercially available Shigella strains? Can I produce one on my own? What happens if I just clone the T7 RNA polymerase gene and transform it to Shigella cells? Does this make sense...?

pDNA

Posted 31 December 2009 - 12:17 AM

Since Shigella is closely related to E. coli it might be possible to use the lysogenization kit from Novagen to make your own DE3-shigella strain.

You can check the sequence ...you need the attB site for integration of the lambda-phage ...otherwise it would not work.

Regards,
p