title: [Expression and activity analysis of catalytic domain of PTP1B].
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 was inserted into vector pET-22b(+) and expressed efficiently in E. coli BL21(DE3) under optimum 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 Ni(2+)-affinity chromatographic column. After purification, the purity of the proteins was more than 95%. Western blotting analysis suggested that the purified proteins could specially combine with anti-PTP1B antibody. Enzyme activity assay showed that the protein has phosphatase activities.
authors.affiliation: Cancer Research Center of Medical College, Xiamen University, Xiamen 361005, China.
authors.forename: Shengyu
authors.initials: S
authors.lastname: Wang
authors.forename: Jianghua
authors.initials: J
authors.lastname: Yan
authors.forename: Yanglin
authors.initials: Y
authors.lastname: Pan
authors.forename: Xuejun
authors.initials: X
authors.lastname: Li
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
mesh_terms: D002384:Catalysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1