Construction of prokaryotic expression vector of human angiogenesis inhibitor arresten and its expression in E.coli
- VernacularTitle:血管生成抑制因子arresten基因原核表达载体的构建及其在大肠杆菌中的表达
- Author:
Zifang SONG
;
Qichang ZHENG
;
Lin ZHU
;
Anbin HU
;
Yiqing LI
;
Xiaogang SHU
;
Yuan TIAN
- Publication Type:Journal Article
- Keywords:
Genes, arresten;
Vectors;
Gene expression
- From:
Chinese Journal of Pathophysiology
1999;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.
