%0 Journal Article
%T Cloning and Expression of Kringle 1-3 Gene of Human Plasminogen and the Purification and Bioactivity of Its Expressed Product<br>人纤溶酶原K1-3基因的克隆、表达和产物的纯化及抗癌活性分析
%A ZHANG Tian-Yuan LUO Jin-Xian
%A LU Xing-Yan
%A <br>张添元
%A 罗进贤
%A 陆幸妍
%J 生物工程学报
%D 2002
%I 
%X Kringle 1-3 domain is a recently found angiogenesis inhibitor with anti-angiogenesis and anti-tumor activity. The kringle 1-3 gene was amplified by PCR technique using angiostatin gene as template. After DNA sequencing, the PCR product was cloned into pPIC9K resulting in recombinant plasmid pPIC9K13 which was then transformed into Pichia pastoris GS115. The high copy integration transformants screened by PCR and G418 methods were cultivated in flasks. The K1-3 was expressed and secreted to the medium and has immunogenic activity as shown by SDS-PAGE and Western blotting. High cell density culture was carried out in 30-liter and 80-liter bioreactor, the biomass reaches 300 OD after methanol induction, and the expressed product is 200 mg/L. The fermentation supernatant was purified by Streamline SP and Phenyl Sepharose Chromatography, the product appears as a single band on SDS-PAGE, with a purity of 95%-96%. The purified product has anti-angiogenesis and anti-tumor activity.
%K K1-3 domain of human plasminogen
%K gene cloning and expression
%K purification
%K anti-tumor activity<br>人纤溶酶原K1－3
%K 表达
%K 抗癌活性分析
%K 基因克隆
%K 纯化
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=3932A165B104BC03&yid=C3ACC247184A22C1&vid=13553B2D12F347E8&iid=94C357A881DFC066&sid=2EAE52BA5B1222A9&eid=6341CCF6B158C5F9&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=4&reference_num=8