%0 Journal Article
%T Cloning of CTB-PROIN Fusion Gene and its Expression in Escherichia coli<br>霍乱毒素B亚单位与胰岛素原融合基因的克隆、表达及重组蛋白特性分析
%A Li Chen
%A Feng-Xiu Ouyang
%A Bing-Jun Qian
%A Hong Ren
%A Qiang Wang
%A Qing-Wu Jiang
%A Yu-Jiong Wang
%A Jing-Bo Liu
%A Wan-Qi Liang
%A Da-Bing Zhang
%A <br>陈丽
%A 欧阳凤秀
%A 钱炳俊
%A 任宏
%A 王强
%A 姜庆五
%A 王玉炯
%A 刘静波
%A 梁婉琪
%A 张大兵
%J 生物工程学报
%D 2005
%I 
%X A fusion gene CTB-PROIN, in which Proinsulin gene was fused to the 3' end of CTB gene by a hinge peptide 'GPGP', was constructed and cloned into pET-30a(+) to obtain a prokaryotic expression vector pETCPI. Subsequently the recombinant plasmid pETCPI was transformed into E. coli stain BL21 (DE3). After induced by IPTG, the expression product was analyzed by sodium dodecyl sulphate-polyacrylamide gel (15%) electrophoresis (SDS-PAGE), and its result indicated that the recombinant protein CTB-PROIN was expressed and accumulated as inclusion bodies. The recombinant CTB-PROIN protein accumulated to the level of 25% of total bacterial proteins. After inclusion bodies was denaturalized and refolded in vitro, significant assembly of monomers had occurred, and the recombinant protein represented assembled pentamers. The results of western blotting analysis also demonstrated that the fusion protein could be recognized by the anti-CT and anti-insulin antibody, respectively. In addition, the result of the CTB-PROIN-GM1 binding assay, that the protein could bind to monosialoganglioside specifically, showed it possesed biological activity in vitro. These results provided the possibility of developing a cheaper and more efficient oral vaccine for type I diabetes using such constructs.
%K Cholera toxin B subunit
%K proinsulin
%K fusion gene CTB-PROIN
%K monosialoganglioside<br>霍乱毒素B亚单位
%K 胰岛素原
%K 融合基因CTBPROIN
%K 神经节苷脂GM1
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=180FF5422BE6FB40&yid=2DD7160C83D0ACED&vid=659D3B06EBF534A7&iid=0B39A22176CE99FB&sid=02DC3A182A5530DF&eid=AA27B676BFCAA4BE&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=31