Expression and activity analysis of catalytic domain of PTP1B / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 553-557, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-342872
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Catalysis
/
Inclusion Bodies
/
Polymerase Chain Reaction
/
Escherichia coli
/
Protein Tyrosine Phosphatase, Non-Receptor Type 1
/
Genetic Vectors
/
Genetics
/
Metabolism
/
Methods
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2008
Type:
Article
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