%0 Journal Article
%T Expression and Identification of Single Chain Antibody of Prokaryotic Expression Vector pOPE101-8E5<br>原核表达载体pOPE101-8E5的改构及单链抗体的表达鉴定
%A XU Jing
%A DIAO Shi-yong
%A ZHANG Lei
%A XU Jie
%A ME Lei
%A <br>许静
%A 刁世勇
%A 张磊
%A 徐洁
%A 孟磊
%J 中国生物工程杂志
%D 2007
%I 
%X In the present study, we inserted the core-streptavidin cDNA into downstream of multi-cloning site of plasmid pOPE101-8E5 by DNA gene recombination technology. And then, the variable fragments of heavy and light chain of the scFv-8E5 were replaced by the scFv-C4 variable fragments to construct the expression vector pOPE101-C4::core-streptavidin. After transformed the vector pOPE101-C4::core-streptavidin into E.coil, the fusion protein C4::core -streptavidin-His-tag can be expressed by inducing with IPTG, and the expression level and activity of the expressed fusion protein analyzed by SDS-PAGE and Western blot. The results show that a scFv-C4::core-streptavidin fusion protein of 45kDa was obtained, which can bind proteins of 60kDa & 45kDa from the KG1a cells lysate simultaneously. The binding function can be detected by the binding of core-streptavidin and biotin directly.
%K Core-Streptavidin Single chain antibody Fusion protein Expression vector<br>链亲和素
%K 单链抗体
%K 融合蛋白
%K 表达载体
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=C7D9A378776DB821&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=E158A972A605785F&sid=13553B2D12F347E8&eid=BC12EA701C895178&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=9