%0 Journal Article
%T Expression and optimization of anti-AFB1 scFv in Escherichia coli<br>抗黄曲霉毒素B1单链抗体在大肠杆菌中的表达及优化
%A Lian Yang
%A Yanhong Zhang
%A Husheng Ding
%A Liyun Wang
%A Wei Chen
%A Hao Zhang
%A <br>杨炼
%A 张艳红
%A 丁虎生
%A 王丽云
%A 陈卫
%A 张灏?
%J 微生物学报
%D 2009
%I 
%X Abstract: Objective] A drawback of the expression of single chain antibody fragment (scFv) in prokaryotic system is the protein accumulation in the cytoplasm as inclusion body. We aimed at high-level production of an anti-aflatoxin B1 scFv in functional form. Methods] The gene of scFv-H4 was cloned into pET22b vector and transformed into E.coli BL21(DE3) and Origami (DE3), respectively. The amount of functional scFv-H4 was optimized in terms of IPTG concentration and induction temperature. Results] scFv-H4 could be expressed in both BL21(DE3) and Origami (DE3). Compared with BL21(DE3), Origami(DE3) could express multifunctional scFv-H4 (35 mg/ml) and less in inclusion body (11% of the total expression). The expression of scFv-H4 was significantly affected by induction temperature rather than IPTG concentration. Conclusion] The pET22b could be used for high-level expression of the functional scFv-H4 in Origami (DE3), which has an oxidative cytoplasm. In addition, the induction at low temperature avoided the formation of inclusion body.
%K Keywords: aflatoxin
%K scFv
%K expression
%K BL21(DE3)
%K pET<br>关键词：黄曲霉毒素
%K 单链抗体
%K 表达
%K BL21(DE3)
%K Origami
%K (DE3)
%K pET22b
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=F28567745B20A92FDB2E08023457E7D3&yid=DE12191FBD62783C&vid=2A3781E88AB1776F&iid=DF92D298D3FF1E6E&sid=750AE535ABE3D62A&eid=68BCD01D0D745EB3&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=17