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High‐level expression of active human plasminogen activator inhibitor type 1 (PAI‐1) in E. coli
Author(s) -
Zhou Aiwu,
Pei Yi,
Wu Haihong,
Dong Xueyin,
Xu Xianxiu
Publication year - 1996
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549600201241
Subject(s) - plasminogen activator , recombinant dna , microbiology and biotechnology , activator (genetics) , urokinase , expression vector , biology , plasminogen activator inhibitor 1 , t plasminogen activator , incubation , chemistry , biochemistry , gene , endocrinology , genetics
Plasminogen activator inhibitor 1 (PAI‐1) is an important regulator of plasminogen activation, which inhibits both tissue‐type plasminogen activator (tPA) and urokinase‐type plasminogen activator (uPA). The DNA sequence encoding mature PAI‐1 protein was inserted into an inducible expression vector. This gene was highly expressed to produce a soluble active protein in E. coli cells. The amount of the recombinant protein was up to 20% of total cellular protein. By efficient purification with a yield of about 15‐20%, the recombinant protein could be purified to homogeneity with its specific activity up to 6.1×104 (uPA) IU/mg. Its inhibitory activity declined during incubation at 37°C with a half life of about 2 hr.