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


CONCLUSIONS
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.
Gong
Xiao-wei
XW
Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Peng
Yi
Y
Liu
Jing-hua
JH
Li
Zhi-jie
ZJ
Deng
Peng
P
Qin
Qing-he
QH
Jiang
Yong
Y
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D003598:Cytoskeletal Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins / Q000235:genetics*
C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D003598:Cytoskeletal Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins