Expression and purification of recombinant N-terminal peptide of human perforin
- VernacularTitle:人穿孔素氨基端肽段的原核表达与纯化
- Author:
Weisong QIN
;
Fangqiu LI
;
Jianguo WU
- Publication Type:Journal Article
- From:
Journal of Medical Postgraduates
2001;14(1):9-11
- CountryChina
- Language:Chinese
-
Abstract:
Objectives: To express and purify N-terminal peptide of human perforin(hPFP-N). Methods: The recombinant expressive plasmid pGEX-KG/hPFP-N was constructed and then introduced into a strain of E. coli BL21(DE3). Upon the induction of IPTG, GST/hPFP-N fusion protein was expressed. The expressed fusion protein was localized in inclusion bodies which could be solubilized by sonication after detergent lauroylsarcosine was added. The fusion protein was purified by affinity chromatography with glutathione agarose. After being cleaved by thrombin, GST and uncleaved fusion protein were removed by glutathione agarose beads once more, then purified recombinant rhPFP-N protein was obtained. Results and Conclusions: GST/hPFP-N fusion protein can be effectively expressed in E. coli and the protein hPFP-N was obtained after the purification process.
