志贺毒素B（ShT-B）亚单位在两种表达载体中表达形式分析

用KpnⅠ和HindⅢ双酶切pGEMTB3克隆质粒,得到大小约为225bp的ShTB基因片段,分别将其插入到经双酶切的pQE40和pQE30表达载体中,构建了2个ShTB的重组表达质粒pQE40B3和pQE30B2,分别转化到E.coliM15,经IPTG诱导后,重组质粒目的蛋白均得到表达。其中,pQE40B3表达蛋白约占菌体总蛋白的37%,主要为包涵体形式。pQE30B2表达蛋白约占菌体总蛋白的16%,主要为可溶性形式,约9.2%。为重组抗原的制备提供了必要的物质基础。

Expression Form Analysis of Shiga Toxin B Subunit Gene in Two Different Expression Vectors

A cloned pGEM TB 3 was digested with Kpn and Hind III double enzymes and obtained 225bp fragment of ShT B (Shiga toxin B). And then the fragment was inserted into expression vectors pQE40 and pQE30 that digested with the double enzymes to construct two ShT B recombinant expression plasmids pQE40ShTB 3 and pQE30ShTB 2 respectively. The two recombinant plasmids were further transferred into E.coli M15, after induced with IPTG, the target proteins of recombinant plasmids are all expressed. Among them expressed protein of pQE40ShTB 3 accounted for 37% of the cell protein, mainly in inclusion body form. And the expressed protein of pQE30ShTB 2 accounted for 16% of cell protein, mainly in soluble form, about 9.2%. Therefore, these have provided necessary substantial foundation for the preparation of recombinant antigen.;