Expression of human BTC with biological activity
- VernacularTitle:人BTC蛋白表达及生物学活性鉴定
- Author:
han-bei, CHEN
;
yan, DONG
;
qing, SU
- Publication Type:Journal Article
- Keywords:
betacellulin;
clone;
expression vector;
purified protein
- From:Journal of Shanghai Jiaotong University(Medical Science)
2006;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.
