High level expression and purification of 1 C-terminal fragment of merozoite surface protein of Plasmodium falciparum in Pichia pastoris
10.3969/j.issn.1002-2694.2005.12.005
- VernacularTitle:恶性疟原虫主要裂殖子表面蛋白1 C末端片段在毕赤酵母表达系统的高效表达与纯化
- Author:
Zhongguang ZHANG
;
Hengmei ZHAO
;
Yuxiang GONG
- Publication Type:Journal Article
- Keywords:
Plasmodium falciparum;
MSP-1;
malaria vaccine;
Pichia pastoris;
expressio
- From:
Chinese Journal of Zoonoses
2005;(12):1047-1051
- CountryChina
- Language:Chinese
-
Abstract:
To obtain an ideal recombinant C-terminal fragment of the merozoite surface protein of Plasmodium falciparum in the Pichia pastoris expression system, the major surface protein-119 (MSP-119) gene sequence bearing the 6-his gene was inserted into expression vector pPIC9k and the target gene was transformed to the susceptible yeast cells GS115 by using electroporation. The multiple inserts were screened and the successfully expressed MSP-119 protein with the relative molecular weight of 12kDa in the supernatants of cell cultures could be detected by SDS-PAGE. Meanwhile, Western blot analysis also demonstrated that this protein reacted with mouse anti-MSP-119 monoclonal antibody, and the expression level of MSP-119 was more than 1.0 g/L. It is concluded that this recombinant protein expressed in the Pichia pastoris expression system resembles the native proteins existed.
