DAB389-Linker-LHRH重组毒素的构建及生物活性的初步测定  

Primary determination for bioactive and construction of DAB389-Linker-LHRH chimeric toxin

作　　者：岳玉环[1] 张国利[1] 吴广谋[1] 何畅[1] 朱平[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《东北师大学报（自然科学版）》2006年第2期118-121,共4页Journal of Northeast Normal University(Natural Science Edition)

基　　金：国家科技部中小企业创新基金资助项目(03C26212200869)

摘　　要：采用基因工程方法,将白喉毒素的毒性部分(DAB389)通过柔性连接肽(Linker)与促黄体激素释放素(LHRH)基因连接起来,克隆到高效表达载体pET28a(+)中,并在大肠杆菌中表达该融合蛋白(DAB389-Linker-LHRH).通过三步层析纯化目的蛋白,获得了纯度达95%的重组毒素DAB389-Linker-LHRH.用MTT法检测其对过度表达LHRH受体的人宫颈癌HeLa细胞的毒性作用,探讨其对HeLa细胞的特异杀伤情况.实验结果显示DAB389-Linker-LHRH对HeLa细胞具有很强的生长抑制作用,其IC50为12.93μg/mL.表明该免疫重组毒素在体外对人宫颈癌HeLa细胞具有明显的杀伤作用,为进一步研制对宫颈癌有特异治疗作用的靶向抗癌药物奠定了基础.By using of genetic engineering techniques, Linked the DAB389 to LHRH with the linker, constructed the expressed vector pDTLL. The chimeric protein DAB389 - Linker - LHRH was constructed sueeessfully and expressed in E. coll BL21（λDE3）. With three purification procedure,the interest protein is about 95 % pure. Cytotoxie of recombinant toxin DAB389 - Linker - LHRH to human carcinoma cells Hela cells growth was analyzed with MTT. Cytotoxieity analysis with MTT showed that the fusion proteins could inhibit the growth of HeLa eells. The IC50 of DAB389 - Linker - LHRH on HeLa cells is 12.93μg/ mL. This indicated that Chimeric toxin DAB389 - Linker- LHRH had obvious cytotoxity to HeLa cells. These finding had established the foundation for the study of targeted drugs.

关 键 词：白喉毒素 促黄体激素释放素 重组毒素 生物活性 

分 类 号：Q784[生物学—分子生物学]