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SHuffle® for hard-to-express proteins

SHuffle cells can be an alternative expression host for proteins that fail to express in common protein expression strains, such as BL21(DE3). This characteristic of SHuffle cells was demonstrated by the expression of a polymerase, which otherwise failed to be expressed in E. coli B and K12 strains. The polymerase lacks any cysteine, and its folding is therefore unrelated to the redox properties of SHuffle cells.

A 67 kD heat-stable polymerase having no cysteines, was expressed at 30ºC from a plasmid under the regulation of T7 promoter in Shuffle K12 (NEB #C3026) vs wild-type E. coli (BL21 or K12). Crude cell lysates were subjected to heat denaturation at 60ºC for 20 min, and separated into total (T) and soluble (S) fractions. Samples were analyzed by SDS-PAGE, and the relative intensity of the bands representing the polymerase is indicated.

Learn about SHuffle® strains for the expression of multi-disulfide bonded and difficult-to-express proteins