%0 Journal Article
%T High-level expression of fusion protein GGH in Pichia pastoris GS115 by constructing a double plasmid co-expression system<br>应用双质粒共表达体系提高融合蛋白GGH在毕赤酵母GS115中的表达量
%A Hui Wang
%A Wenfang Dou
%A Xiaomei Zhang
%A Hongyu Xu
%A Zhenghong Xu
%A <br>王慧
%A 窦文芳
%A 张晓梅
%A 许泓瑜
%A 许正宏
%J 生物工程学报
%D 2011
%I 
%X In order to make a large-scale preparation of(GLP-1A2G)2-HAS (GGH), the double-plamid pPICZalphaB and pPIC9K co-expression system was introduced into Pichia pastoris GS115. Firstly, the GGH fusion gene was amplified by PCR to create the recombinant expression plasmid pPICZalphaB-ggh, which was transformed into P. pastoris GS 115/F2 that was integrated by another recombinant expression plasmid pPIC9K-ggh. The immunology method combined with high concentration antibiotic was used to screen recombinant strain P. pastoris GS115/F3 capable of high-level expression of GGH protein. The GGH fusion protein expressed by GS115/F3 increased 49.7% compared with the GS 115/F2 in the expression conditions of 3% methanol inducing 80 h at 30 degrees C. At the same time, the quantitative PCR results showed that GGH gene dose in GS115/F3 increased 26.7% with respect to that of GS 115/F2. Furthermore, the Western blotting experiment indicated that the recombinant GGH possess the two antigenicities of GLP-1 and HSA.
%K fusion protein GGH
%K double plasmid
%K Pichia pastoris
%K quantitative PCR
%K Western blotting<br>融合蛋白GGH，双质粒，毕赤酵母，荧光定量PCR，Western
%K blotting
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=08FC36D46A3865B2C1AF797FEAB71651&yid=9377ED8094509821&vid=DB817633AA4F79B9&iid=DF92D298D3FF1E6E&sid=88B4027FEBE4F5FF&eid=A73A882009D0AEFE&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=13