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Protein expression with T7 promoter

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#1 Spright



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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...?

#2 pDNA



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Posted 31 December 2009 - 12:17 AM

Since Shigella is closely related to E. coli it might be possible to use the [url="http://www.merck-chemicals.de/life-science-research/lambda-de3-lysogenization-kit-plus-plyss-und-plyse/EMD_BIO-69725/p_T5yb.s1OKwIAAAEj4Bt9.zLX"]lysogenization kit [/url]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.


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