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Journal of Biomedical Engineering ; (6): 820-824, 2009.
Article in Chinese | WPRIM | ID: wpr-294561

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).


Subject(s)
Animals , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Extracellular Space , Genes, Protozoan , Leishmania donovani , Genetics , Plasmids , Genetics , Protozoan Proteins , Genetics , Recombinant Fusion Proteins , Genetics
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