小麦组蛋白H1基因保守序列原核载体的构建及表达  

Construction and Expression of Prokaryotic Expression Vector of Wheat Histone H1 Gene Conservative Sequence

作　　者：宋红肖[1] 谭广云[1] 王春艳[1] 张瑞兴[1] 朱乾琨[1] 孙浩然[1] 

机构地区：[1]吉林大学农学部畜牧兽医学院,长春130062

出　　处：《吉林农业大学学报》2010年第1期39-42,共4页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目(30170694)

摘　　要：采用PCR技术扩增TAT-H1bs融合基因,并通过基因操作构建了融合基因的原核重组载体pET-28a-TAT-Hlbs。将阳性重组质粒转化至受体菌Rosseta(DE3)感受态细胞中,以IPTG诱导表达,表达产物经聚丙烯酰胺凝胶电泳检测和Western-blot检测。结果表明:重组菌能够表达目的蛋白,软件分析结果表明表达蛋白约占菌体蛋白的40%,上清表达量约为20%。上清蛋白经纯化后,Western-blot结果显示,His-Tag单克隆抗体可以很好地与所表达的蛋白带特异性结合。所获得的融合蛋白以高效胞质可溶形式表达。TAT-Hlbs fusion gene was obtained through PCR amplification, and recombinant expression plasmid pET-28a-TAT-Hlbs was constructed by gene operation technology. The correct recombinant expression plasmid was transformed into the host strain Rosseta induced by IPTG. The specific protein expressed was detected by SDS- PAGE and Western blot. The result showed that the targeted protein was detected in Rosscta（DE3）, the fusion protein was expressed at high level amounting to 40% of the total bacterial protein analyzed by computer software. The amount in supernatant is about 20% . After the supernatant protein was purified, Western blot showed the monoclonal antibody anti-His-Tag could react to the targeted protein expressed specifically. The fusion protein was expressed highly in cytoplasmic fraction in Rosscta（DE3） .

关 键 词：小麦组蛋白H1 融合蛋白 可溶性表达 穿膜肽TAT 

分 类 号：Q786[生物学—分子生物学]