Cloning, sequencing of human angiostatin gene and its expression in E. coli
自然科学进展(英文版)(1999)
摘要
Angiostatin is a recently discovered angiogenesis inhibition factor which can inhibit the growth of tumor and its metastasis and has perspective for clinical application. Human angiostatin gene was amplified by polvmerase chain reaction using human plasminogen cDNA as template based on the fact that angiostatin is an internal fragment of plasminogen. The amplified fragment was sequenced and cloned in plasmid pBV220 resulting in plasrmd pBV N2 winch was then transformed into E. coli DH5a. Angiostatin was efficiently expressed in E. coli by temperature induetren SDS-PAGE and Western blot analyses indicate that the expressed product constitutes 38% of the total bacterial proteins, equivalent to 212 mg/L fermentation broth, and reacts with antiserum against human plasminogen. The recombinant human angiostatin inhibits basic fibroblast growth factor induced angiogenesis on chick embryo chorioallantoic membrane and growth of primary B16 melanoma in C57BL/6J mouse.
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关键词
human angiostatin gene,cloning
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