%0 Journal Article
%T Secretion Expression of Mt. Heat Shock Protein 70 in Pichia pastoris and Identification of the Protein<br>结核杆菌热休克蛋白70在毕赤酵母中的分泌表达与鉴定
%A PENG Ming
%A Li
%A LING Ning
%A XU Hong
%A Mei
%A QING Yu
%A Ling
%A REN Hong
%A <br>彭明利
%A 凌宁
%A 许红梅
%A 卿玉玲
%A 任红
%J 生物工程学报
%D 2003
%I 
%X To obtain the expression of Mycobacterium tuberculosis heat shock protein 70 in methylotropic yeast. The expression vector pPIC9K-hsp70 was constructed, linearized and introduced into Pichia pastoris GS115 by electroporation. The result protein was secreted into the supernatant induced by 0.5% methanol at 30 degrees C and purified by centrifugation, ultrafiltration and ATP-agarose. The recombinant Hsp70 was identified by SDS-PAGE, Western blot, mice experiment and effect on the immature DC. The SDS-PAGE and Western blot analysis showed that the apparent molecular weight of expressed Hsp70 was about 70 kD and the expressed protein could specifically react with anti-Mt. Hsp70 IgG. And mice immunization indicated the expressed hsp70 had immunogenicity. Hsp70 could induce DC maturation and release Th1 cytokine. The secreted 70 kD protein was about 120 mg/L which accounted for more than 30% of the total supernatant protein and was purified to electrophoretic purity. The Hsp70, which had the biological activity, is successfully secretorily expressed in the Pichia pastoris GS115.
%K heat shock protein 70
%K Pichia pastoris
%K secretion expression
%K dendrite cell<br>结核杆菌热休克蛋白70
%K Pichia
%K 
%K 分泌表达
%K 树突状细胞
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=F341DF117C67ADD3&yid=D43C4A19B2EE3C0A&vid=2A8D03AD8076A2E3&iid=38B194292C032A66&sid=11CEECA6DA9E4AC5&eid=211C7E02AC474301&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=4&reference_num=8