凋亡素融合基因原核表达载体的构建及可溶性表达  

Construction of Apoptin Fusion Gene Prokaryotic Expression Vector and Its Soluble Expression

作　　者：王春艳[1] 郝军元[1] 张英波[1] 王诺[1] 张玉静[1] 

机构地区：[1]吉林大学人兽共患病研究所,长春130062

出　　处：《吉林农业大学学报》2007年第3期271-274,278,共5页Journal of Jilin Agricultural University

基　　金：国家自然科学基金资助项目(30170694)

摘　　要：在获得凋亡蛋白融合基因的基础上,成功地构建了重组表达质粒PET-22b-SPA,将阳性重组质粒转化表达受体菌BL21(DE3)感受态细胞,经IPTG诱导表达,表达产物经聚丙烯酰胺凝胶电泳检测,凋亡蛋白融合基因获得高效表达。软件分析结果表明表达蛋白占菌体蛋白的20%,上清表达量约为10%。上清蛋白经纯化后,Western blot结果显示,利用凋亡蛋白单克隆抗体可以很好地与所表达的蛋白带特异性结合。Recombinant expression plasmid pET-22b-SPA was constructed according to recombinant plasmid.The correct recombinant expression plasmid was transformed into the host .strains BL21 （DE3） induced by IPTG. The specific expressed protein was detected by SDS-PAGE . The recombinant protein was expressed at high level accounting for 20 % of the total bacterial protein analyzed by computer software. The amount in supematant is about 10%. After the supernatant protein was purified, western blot showed the monoelonal antibody raised against the recombinant apoptin in murines could react to the protein expressed specifically.

关 键 词：凋亡蛋白 融合基因 可溶性表达 

分 类 号：Q786[生物学—分子生物学]