Construction of recombinant plasmids and their overexpression of SIK2 cDNA and its truncated mutants
10.7652/jdyxb201403002
- VernacularTitle:SIK2 cDNA及其截短体原核表达重组体的构建及表达
- Author:
Jianxiao WANG
;
Shanshan LI
;
Xu LIU
;
Pan WANG
;
Xiaodan LIU
;
Yu WANG
;
Shanshan ZHAO
;
Pingkun ZHOU
;
Yongqing GU
- Publication Type:Journal Article
- Keywords:
salt-inducible kinase;
SIK2;
GST;
fusion protein;
induced expression;
recombinant plasmid
- From:Journal of Xi'an Jiaotong University(Medical Sciences)
2014;(3):290-294
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct recombinant plasmids of SIK2 cDNA and its truncated mutants and induce its expression in E.coil.Methods We designed primers of SIK2 and its truncated mutants.The gene fragments of SIK2,SIK2-Δ1 (280-926),SIK2-Δ2 (400-926),SIK2-Δ3 (1-400),and SIK2-Δ4 (700-926)were amplified by polymerase chain reaction (PCR)and cloned into pGEX-4T-2 vector to construct recombinant plasmids with GST. The plasmids were transformed into E.coil BL2 1 respectively,and induced with IPTG to express fusion protein. The results were confirmed by Coomassie blue staining and Western blot.Results We successfully constructed recombinant plasmid of SIK2 cDNA and its truncated mutants.Coomassie blue staining and Western blot resutls showed that these plasmids were induced to be expressed in E.coil BL21.Conclusion SIK2 cDNA and its truncated mutants were overexpressed in E.coil BL2 1 ,which lays expereimental foundation for further study on the function of each domain of SIK2 .
