抗阿尔茨海默病神经保护肽humanin基因的克隆与表达  

The cloning and expression of an anti-Alzheimer's disease neuroprotective peptide gene humanin

作　　者：罗本燕[1] 袁敏[1] 唐敏[1] 陈智[2] 

机构地区：[1]浙江大学医学院附属第一医院神经内科,杭州310003 [2]浙江大学医学院传染病研究所,杭州310003

出　　处：《中华神经科杂志》2004年第3期231-233,共3页Chinese Journal of Neurology

基　　金：浙江省科委基金资助项目 [2 0 0 3 (3 0 0 0 40 ) ]

摘　　要：目的　利用基因工程技术克隆和表达humanin基因。方法　人工合成humanin基因片段并克隆到原核表达载体pGEMEX 1和pet4 4a中。再将此等质粒转化至大肠杆菌 ,经过筛选鉴定获得高表达阳性克隆。结果　基因序列分析显示 ,humanin基因片段被成功地克隆到原核载体中 ;聚丙烯酰胺凝胶电泳 (SDS PAGE)显示 ,在异丙基硫代 β D 半乳糖苷 (IPTG)的诱导下 ,pGEMEX 1 humanin表达的融合蛋白的相对分子质量约为 2 80 0 0 ,融合蛋白含量占细菌表达蛋白总量的 4 0 % ;而pet4 4a humanin表达的融合蛋白的相对分子质量约为 6 30 0 0 ,融合蛋白表达量占细菌蛋白总量的 2 0 %。结论　利用基因工程技术可成功地克隆和表达humanin基因 ,并可获得高效表达。Objective To use gene engineering technique to express the humanin.Methods Humanin segment was synthesized and cloned into the prokaryocyte expression vector pGEMEX-1 and pet44a. The plasmid was transformed to E. coli BL21(DE3) by screening and identifying to obtain the high expressing positive clone.Results The humanin segment was cloned into vector successfully,and expressed efficiently with the inducement of IPTG. The molecular weight of the expressed recombinant protein of pGEMEX-1-humanin was about 28 kd while it was 63 kd in pet44a-humanin,and the expressed quantity of fusion protein was 40 percentage of the total bacterial protein while it was 20 percent in pet44a-humanin.Conclusion The humanin has been cloned and expressed successfully and the high efficiently expressed recombinant protein might contain the intact important structure domain of the humanin.

关 键 词：蛋白质类 重组融合蛋白质类 阿尔茨海默病 分子克隆 基因表达 

分 类 号：R749.16[医药卫生—神经病学与精神病学]