Expression and activity analysis of catalytic domain of PTP1B.
- Author:
Shengyu WANG
1
;
Jianghua YAN
;
Yanglin PAN
;
Xuejun LI
;
Zhong CHEN
Author Information
1. Cancer Research Center of Medical College, Xiamen University, Xiamen 361005, China.
- Publication Type:Journal Article
- MeSH:
Catalysis;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Humans;
Inclusion Bodies;
metabolism;
Polymerase Chain Reaction;
methods;
Protein Tyrosine Phosphatase, Non-Receptor Type 1;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2008;24(4):553-557
- CountryChina
- Language:Chinese
-
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.
