[Cloning and expression of extracellular region gene located in N-terminus of Leishmania Donovani].
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).
Chen
Xian
X
Department of Parasitology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Chen
Jianping
J
Xu
Jia'nan
J
Wang
Xin
X
Lu
Rui
R
Lu
Dianxiang
D
Ho
Xiaosu
X
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005110:Extracellular Space; D017125:Genes, Protozoan; D007893:Leishmania donovani / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins