Cloning and expression of extracellular region gene located in N-terminus of Leishmania Donovani.
- Author:
Xian CHEN
1
;
Jianping CHEN
;
Jia'nan XU
;
Xin WANG
;
Rui LU
;
Dianxiang LU
;
Xiaosu HO
Author Information
1. Department of Parasitology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Extracellular Space;
Genes, Protozoan;
Leishmania donovani;
genetics;
Plasmids;
genetics;
Protozoan Proteins;
biosynthesis;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics
- From:
Journal of Biomedical Engineering
2009;26(4):820-824
- CountryChina
- Language:Chinese
-
Abstract:
The objective of this study was to construct and express recombinant prokaryotic plasmid pET32a (+)- ast1 in E. coli BL21(DE3). Amastin gene was amplified from genomic DNA of Leishmania Donovani and its transmembran region was predicted by the methods of SOSUI and Tmpred; astl located in N-terminus of amastin gene was amplified and cloned into prokaryotic plasmid pET32a(+), which was named pET32a(+)-ast1, and then rAST1 was expressed in E. coli BL21(DE3). The results of SDS-PAGE and immunobloting assay showed that a fusion protein rAST1 (relative molecular mass about 27 kDa) was able to express in BL21. The recombinant prokaryotic plasmid pET32a(+)- ast1 was successfully constructed, and noted to be efficiently expressed in E. coli BL21(DE3).
