人微小病毒3个主要蛋白基因真核表达载体的构建  

Construction of Eukaryotic Expression Plasmids Containing Gene of 3 Major Proteins of Parvovirus B19

作　　者：张伟平[1] 聂占国[1] 代忠明[1] 热合曼[1] 游云[1] 雷婷[1] 陈红[2] 张新[1] 刘国伟[1] 

机构地区：[1]兰州军区乌鲁木齐总医院消化病中心,新疆乌鲁木齐830000 [2]兰州军区乌鲁木齐总医院输血科,新疆乌鲁木齐830000

出　　处：《现代生物医学进展》2012年第31期6019-6021,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助项目(30873472);中国博士后科学基金资助项目(20090461485)

摘　　要：目的:分别克隆人细小病毒B19三个主要蛋白VP1、VP2、NS1全长基因,构建真核表达载体。方法:利用PCR和分子克隆技术,分别将B19病毒vp1、vp2、ns1基因全长片段扩增后,构建带荧光标签的真核表达载体;在人体细胞中表达并通过荧光、RT-PCR和Western Blot、测序等方法鉴定。结果:成功构建了包含B19病毒vp1、vp2、ns1全长基因,并在人体细胞中表达了VP1、VP2、NS1蛋白。结论:人微小病毒B19三个主要蛋白基因得到克隆和表达,为进行相关的研究奠定了基础。Objective： To construct eukaryotic expression plasmids containing gene of 3 major proteins of parvovirus B19. Methods： vp1, vp2 and nsl genes of parvovirus B19 were cloned and were constructed into pReceiver-M03, pReceiver-M33 and pReceiver-M16 plasmid. RT-PCR, fluorescence microscope and Western Blot assays were used to identify these plasmids. Results： After transfection, vpl, vp2 and nsl nucleotide acids and proteins were found in target cells. Conclusion： Three eukaryotic expression plasmids containing gene of 3 major proteins ofparvovirus B19 were constructed.

关 键 词：微小病毒B19 真核表达 质粒 

分 类 号：Q75[生物学—分子生物学] Q78