SHuffle Expression

The SHuffle® suite of competent E. coli was designed to encourage disulfide bond formation in expressed proteins. Additionally, tight control of expression by lacIq allows for the expression of potentially toxic genes. The SHuffle suite includes SHuffle Express Competent E. coli (NEB #C3028) SHuffle T7 Competent E. coli (NEB #C3026), SHuffle T7 Express lysY Competent E. coli (NEB #C3030) and SHuffle T7 Express Competent E. coli (NEB #C3029).


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FAQs for SHuffle Expression
Protocols for SHuffle Expression
Application Notes for SHuffle Expression
PfCHT1 Chitinase Expression in Shuffle® T7 Express
Plasmodium falciparum chitinase (PfCHT1) with three cysteines was expressed from a plasmid under the regulation of T7 promoter. After induction, cells were harvested and crude cell lysates were prepared. PfCHT1 was assayed using a chromogenic substrate (CalBioChem #474550) and standardized to protein concentration using Bradford reagent.
vtPA Activity Expression in Shuffle®
Truncated tissue plasminogen activator (vtPA), which contains nine disulfide bonds when folded and oxidized correctly, was expressed from a pTrc99a plasmid in the cytoplasm of E. coli cells. After induction, cells were harvested and crude cell lysates were prepared. vtPA was assayed using a chromogenic substrate Chromozym t-PA (Roche #11093037001) and standardized to protein concentration using Bradford reagent. E. coli wt + cells are DHB4, which is the parent of FÅ113 (Origami™).
Disulfide Bond Formation
Disulfide bond formation in the cytoplasm of wild type E. coli is not favorable, while SHuffle is capable of correctly folding proteins with multiple disulfide bonds in the cytoplasm.
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