Article ID Journal Published Year Pages File Type
2079004 Chinese Journal of Biotechnology 2008 5 Pages PDF
Abstract
The amino acid sequence (1-301aa) coding the human PTP1B catalytic domain (PTP1Bc) was obtained from the GenBank. The PTP1Bc gene was constructed by overlapping PCR, then inserted into vector pET-22b (+) and expressed efficiently in E. coli BL21 (DE3) under the optimization condition after IPTG induction. The proteins were expressed mainly as inclusion bodies with the yield of more than 30% of total bacterial proteins. The expressed products were purified through Ni2+-affinity chromatographic column. After purification, the purity of the proteins was more than 95%. The result of western blotting confirmed that the purified proteins could specially combine with anti-PTP1B antibody. The enzyme activities experiment showed that the protein had phosphatase activities. The gene construction, expression, and purification of PTP1Bc may provide basis for further study of its functions.
Related Topics
Life Sciences Biochemistry, Genetics and Molecular Biology Biotechnology
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