α-银环蛇毒素同工毒素基因的克隆和表达  

Molecular Cloning and Expression of an Isotoxin Gene,α-bungarotoxin,from Bungarus multicinctus

作　　者：汪芳[1] 王义权[1] 

机构地区：[1]厦门大学生命科学学院

出　　处：《Acta Genetica Sinica》2005年第7期682-688,共7页

基　　金：国家自然科学基金(编号:30470938);教育部骨干教师资助计划项目(编号:GG180210024031740);教育部留学回国人员启动基金资助~~

摘　　要：α银环蛇毒素是存在于银环蛇蛇毒中的一种长链神经毒素,能与神经递质受体特异性结合,因此它不仅是神经信号传导机制研究的重要分子探针,更具有开发治疗某些神经性疾病新型药物的可能性。用SMART技术,反转录合成银环蛇毒腺cDNA,克隆并测定了α银环蛇毒素基因,得到14种mRNA序列;进而将其中一种新的α银环蛇毒素基因亚克隆到原核表达载体pQE30a和pGEX4T1中,在大肠杆菌中通过IPTG诱导表达,其中带有GST融合蛋白的重组质粒αBgTX(P22A31)/pGEX4T1在BL21菌株中得到高效表达,表达量占菌总蛋白的30%,可溶形式存在的融合蛋白大于25%。Snake venom contains a number of small proteins,enzymes and other components,which displays a broad spectrum of biological activities.With the ability of specifically binding on acetylcholine acceptor,α-bungarotoxins are not only useful molecular probes in investigating the mechanism of neural signal transmission,but also potential pharmic preparations for neural disease treatment.In current research,cDNAs of Bungarus multicinutus venom gland were synthesized using SMART^TM cDNA amplification kit and then,α-bungarotoxin genes were cloned and sequenced.Total of 20 clones were sequenced representing 14 isotoxin mRNAs of α-bungarotoxins.Among those clones,a novel isotoxin gene was subcloned into two expression plasmids,α-BgTX/pQE30a and α-BgTX/pGEX-4T-1,and transformed into E.coli.After inducing with IPTG,fused protein of GST-α-BgTX was successfully expressed at level of 30% gross proteins of bacteria.More than 25% of fused protein was in the soluble fraction and the rest in inclusion body.

关 键 词：Α-银环蛇毒素 基因克隆 原核表达 CDNA序列 

分 类 号：Q78[生物学—分子生物学]