P1因子分子克隆的体外感染性分析  

Infectivity Analysis of Cloned Genomic DNA of P1 Agent in vitro

作　　者：温立斌[1] 何孔旺[1] 杨汉春[2] 

机构地区：[1]江苏省农业科学院兽医研究所,南京210014 [2]中国农业大学动物医学院,北京100193

出　　处：《畜牧兽医学报》2008年第7期941-944,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家“973”计划前期研究专项(2007CB116308);江苏省博士后基金(5910602);江苏省农业科学院博士后基金(6510501)

摘　　要：利用脂质体分别将类猪圆环病毒2型的P1因子的单拷贝分子克隆和双拷贝串联分子克隆转染PK-15细胞，电镜检测可在转染P1因子双拷贝串联分子克隆的PK-15细胞的胞浆和胞核中发现包涵体。相比较而言，胞浆包涵体数量较多，基本呈圆形，直径0.1～0.3 μm；胞核包涵体有2种类型：一种为小而圆的、直径约0.1 μm的高电子密度的小体；一种是大小为0.4～0.8 μm、略呈六角形的包涵体。将转染细胞连续传代5次，通过PCR可在转染P1因子双拷贝串联分子克隆的细胞传代物中检测到P1因子的DNA。这是首次报道P1因子的分子克隆在体外具有感染性，为进一步研究P1因子在体内的生物学意义奠定了物质基础。The ultrastructure of porcine kidney （PK）-15 cells was examined after lipofectamine transfection of the molecular clone of the P1 agent. PK-15 cells transfected with the tandem dimmer of the P1 molecular DNA clone had numbers of intracytoplasmic inclusions, and a few cells had intranuclear inclusions. Intracytoplasmic inclusions were round to oval and 0.1-0.3μm in diameter, and intranuclear inclusions were more electron dense than intracytoplasmic inclusions and were of two general types： The first type were round and small （0.1 μm approximately） and the second type were hexagon and larger （0.4-0. 8 μm in diameter）. The cells transfected with the tandem dimmer of the P1 molecular DNA clone were positive for P1 DNA at passages 5. This is the first report that the P1 molecular clone has infectivity in vitro and it will provide fundamental materials for further study on the biological characterization of P1.

关 键 词：P1因子 分子克隆 体外 感染性分析 

分 类 号：S852.659.2[农业科学—基础兽医学]