New ligation independent cloning vectors for expression of recombinant proteins with a self-cleaving CPD/6xHis-tag.

ABSTRACT:

Background

Recombinant protein purification is a crucial step for biochemistry and structural biology fields. Rapid robust purification methods utilize various peptide or protein tags fused to the target protein for affinity purification using corresponding matrices and to enhance solubility. However, affinity/solubility-tags often need to be removed in order to conduct functional and structural studies, adding complexities to purification protocols.

Results

In this work, the Vibrio cholerae MARTX toxin Cysteine Protease Domain (CPD) was inserted in a ligation-independent cloning (LIC) vector to create a C-terminal 6xHis-tagged inducible autoprocessing enzyme tag, called "the CPD-tag". The pCPD and alternative pCPD/ccdB cloning vectors allow for easy insertion of DNA and exp

SUBMITTER: Biancucci M 

PROVIDER: S-EPMC5216533 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature