鸡贫血病毒全长基因感染性分子克隆的构建  

Construction of Infectious Molecular Clone of the Full-Length DNA from CAV

作　　者：李秀梅[1] 江丽萍[2] 郭立力[1] 李颖[1] 朱雅宁[1] 石瑜[1] 杨爱华[1] 陈荣荣[1] 

机构地区：[1]天津市动物疫病预防控制中心,天津300402 [2]山西农业大学,山西太谷030801

出　　处：《动物医学进展》2011年第8期6-11,共6页Progress In Veterinary Medicine

基　　金：天津市应用基础及前沿技术研究计划基础项目(08JCYBJC04700)

摘　　要：PCR法扩增鸡贫血病毒全长基因组,将2个全长基因组顺向连接插入到pBluescriptΠSK(+)质粒载体中,获得Psk2CAV质粒并转染MDCC-MSB1细胞,盲传4代后,用间接免疫荧光抗体试验(IFA)能检测到特异性荧光。将Psk2CAV质粒DNA于腿部肌肉接种10羽1日龄CAV阴性SPF鸡。剖检结果显示,阳性对照组和Psk2CAV质粒组均有鸡只出现腿肌、胸肌轻微出血,胸腺出现萎缩或出血。组织病理学结果显示,阳性对照组和Psk2CAV质粒组胸腺小叶萎缩,出血、淤血,髓质部萎缩,淋巴细胞减少。通过PCR可以分别从体外感染MDCC-MSB1细胞和体内感染鸡只中扩增到病毒基因片段。本研究证明含有双拷贝CAV的重组质粒Psk2CAV在体内、外均具有感染性,能衍生出病毒,并产生符合自然感染CAV的大体病变和组织病理学变化。The full-length genome of CAV were amplified by PCR, and ligating two full-length genome of CAV in tandem into pBluescript II SK（＋） vector, named as Psk2CAV , was transfected into MDCC-MSB1 cell, which IFA test showed specificity fluorescence. Then, ten chicks were intramuscularly inoculated with 100 μg of the coined CAV plasmid. The result from necropsy illustrated that showed slight bleeding in crureus and pectorals,and atrophy or bleeding in thymusin in positive control group and Psk2CAV plasmid group. The result of pathological section demonstrated that lobule of thymus has atrophy, bleeding and gore, medulla has atrophy, and lymphocyte has a lessening. Genome of CAV were amplified from infected MDCC-MSB1 cell in vivo and infected chick in vitro. The results indicate that the cloned CAV genomic DNA is infectious in vivo and in vitro, and the derived virus could replicate the basic same pathological change and pathological change of CAV of natural infection.

关 键 词：鸡贫血病毒 聚合酶链反应 感染性分子克隆 转染 

分 类 号：S852.659.2[农业科学—基础兽医学] Q785[农业科学—兽医学]