Construction and expression of the fusion vector of His-tagged human ARPC2 gene] |
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Authors: | Xiao-wei Gong Yi Peng Jing-hua Liu Zhi-jie Li Peng Deng Qing-he Qin Yong Jiang |
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Institution: | Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China. |
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Abstract: | OBJECTIVE: To construct the expression vector of His-ARPC2 fusion protein and obtain its expression and purification 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. |
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