Cloning and expression of mouse canstatin cDNA in E.coli
- VernacularTitle:小鼠canstatin cDNA的克隆及其在大肠杆菌中的表达
- Author:
Weihong HOU
;
Baomei YUAN
;
Tianyun WANG
;
Yurong CHAI
;
Guiqin HOU
;
Jianmin WANG
;
Lexun XUE
- Publication Type:Journal Article
- Keywords:
Canstatin;
cDNA cloning;
Angiogenesis inhibitors;
Prokaryotic expression
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To clone and express mouse canstatin (m canstatin)cDNA and provide a basis for the further research on its anti-angiogenic activity and potential application for cancer therapy. METHODS: Total RNA was extracted from mouse liver tissue by Trizol Reagent, and mouse canstatin cDNA was amplified by RT- PCR, then cloned into vector pMD18-T for sequencing. pET30a(+)-m canstatin recombinant plasmid was constructed and expressed in E.coli BL21 with induction of IPTG. RESULTS: Mouse canstatin cDNA is 684 bp coding 227 amino acids. The sequences of both cDNA and amino acid share high homology with human canstatin, with cDNA identity at 89% and amino acids identity at 96% to human canstatin. In the present study, pET30a(+)-m canstatin recombinant plasmid was expressed in E.coli BL21. CONCLUSION: Mouse canstatin cDNA has been cloned for the first time. Constructed pET30a(+)-m canstatin recombinant plasmid is highly expressed in E.coli BL21.
