Construction and expression of the fusion vector of His-tagged human ARPC2 gene

Abstract

Abstract: Objective To construct the expression vector of His-ARPC2 fusion protein and obtain its expression and purifica-tion in E.coli. Methods ARPC2 cDNA codon region was amplified by PCR from human liver cDNA library and cloned into pET-14b vector following the routine procedures. After identification by enzyme digestion, PCR and sequencing, the positive clones were transformed into BL21 (DE3) competent cells, and the expression of His-ARPC2 fusion protein was induced with IPTG and further purified by Ni-NTA affinity chromatography. Results The constructed His-ARPC2 fusion protein vector was highly efficiently expressed in E.coli. With Ni-NTA affinity chromatography, a purified His fusion protein with relative molecular mass of approximately 36 000 was obtained. Conclusion The expression vector of His-ARPC2 fusion protein is constructed, expressed and purified under non-denaturing conditions, which may significantly facilitate future study of the physiological functions of ARPC2 and characterization of its interaction proteins.

CLC Number:

GONG Xiao-wei, PENG Yi, LIU Jing-hua, LI Zhi-jie, DENG Peng, QIN Qing-he, JIANG Yong. Construction and expression of the fusion vector of His-tagged human ARPC2 gene[J]. Journal of Southern Medical University, 2004, 24(06): 628-630,635.

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