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Segmental expression and C-terminal labeling of protein ERp44 through protein trans-splicing.

Protein Expr Purif. 2015; 
Dai X, Liu XQ, Meng Q.
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Monoclonal Antibody Services … electrophoresis in SDS–polyacrylamide gel. Western blotting was carried out using anti-Flag (DYKDDDD) monoclonal antibody (GenScript) and Enhanced Chem- luminescence detection kit (GE Healthcare). Intensities of protein bands … Get A Quote

摘要

Endoplasmic reticulum resident protein 44 (ERp44) is a member of the protein disulfide isomerase family and functions in oxidative protein folding in the endoplasmic reticulum. A structurally flexible C-terminal tail (C-tail) of ERp44 plays critical roles in dynamically regulating ERp44's function in protein folding quality control. The structure-function dynamics of ERp44's C-tail may be studied further using fluorescence and other techniques, if methods are found to label the C-tail site-specifically with a fluorescent group or segmentally with other desired labels. Here we have developed such methods, employing split inteins capable of protein trans-splicing, and identifying atypical S1 split inteins able to... More

关键词

C-terminal labeling; ERp44; Intein; Protein trans-splicing; Segmental expression