透明颤菌血红蛋白基因在生黑葡萄糖酸杆菌中的克隆与表达  

Cloning and expression of Vitreoscilla Hemoglobin gene in Gluconobacter melanlgenu

作　　者：陈羽[1] 徐威[1] 刘华松[1] 位星[1] 张世野[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2016年第7期577-580,共4页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省教育厅项目(L2013380);辽宁省教育厅高等学校创新团队项目(LT201423)

摘　　要：目的在生黑葡萄糖酸杆菌中克隆并表达透明颤菌血红蛋白基因(Vitreoscilla Hemoglobin gene,vgb)。方法构建重组质粒p UC19-vgb并电转化生黑葡萄糖酸杆菌。采用十二烷基硫酸钠(sodium dodecyl sulfonate,SDS)-聚丙烯酰胺凝胶电泳和一氧化碳(carbon monoxide,CO)-示差光谱进行血红蛋白表达和生物活性测定。结果 SDS-聚丙烯酰胺凝胶电泳显示有大量与目的蛋白大小相当(约15 ku)的蛋白表达。CO-示差光谱在420 nm处有一吸收峰,显示重组质粒p UC19-vgb成功转化入生黑葡萄糖酸杆菌,并表达具有生物活性的透明颤菌血红蛋白(Vitreoscilla Hemoglobin,VHb)。结论成功构建vgb重组质粒,并实现在生黑葡萄糖酸杆菌中表达VHb。Objective To clone and express Vitreoscilla Hemoglobin gene（ vgb） in G luconobacter melanlgenu. Methods Recombinant plasmid p UC19-vgb was constructed and introduced into G luconobacter melanlgenu. The expression of VHb was verified by SDS-PAGE method and the CO-binding difference spectra. Results The expression of VHb protein with molecular weight of 15 ku was observed by the SDSPAGE. The activity of expressed VHb was detected by CO-binding assay,and the peak at 420 nm has confirmed the positive activity,which indicated that recombinant plasmid p UC19-vgb was introduced into G luconobacter melanlgenu successfully. Conclusions Recombinant plasmid vector p UC19-vgb was successfully constructed and expressed in G luconobacter melanlgenu.

关 键 词：生黑葡萄糖酸杆菌 透明颤菌血红蛋白 转化 

分 类 号：R786[医药卫生—口腔医学]