传染性法氏囊病病毒cDNA文库的构建  

CONSTRUCTION OF IBDV cDNA LIBRARY FROM DOUBLE STRANDED RNA OF INFECTIOUSBURSAL DISEASE VIRUS

作　　者：陈士友[1] 张兹钧 金由辛 陈溥言[1] 蔡宝祥[1] 王德宝 

机构地区：[1]南京农业大学动物医学系,珠海拱北动植物检疫局,上海中科院上海生物化学研究所分子生物学国家重点实验室,上海中科院上海细胞生物学研究所

出　　处：《病毒学报》1994年第2期159-163,共5页Chinese Journal of Virology

基　　金："863"高科技项目;高校博士学科点专项;上海分子生物学国家重点实验室

摘　　要：本试验利用随机引物法反转录鸡传染性法氏囊病病毒（Ｉｎｆｅｃｔｉｏｕｓｂｕｒｓａｌｄｉｓｅａｓｅｖｉ－ｒｕｓ，ＩＢＤｖ）的双链ＲＮＡ（ｄｓＲＮＡ），并构建了ｃＤＮＡ文库。病毒为青岛株，来自感染鸡的法氏羹。用免疫复合物沉淀法提纯病毒，抽提核酸，经琼脂糖凝胶电泳纯化。ＩＢＤＶｄｓＲＮＡ经反转录、加ｐｏｌｙ（ｄｃ）后，克隆于加有ｐｏｌｙ（ｄＧ）的线性化质粒ｐＴ／３α－１９上，并转化大肠杆菌ＤＨ５αＦ′，结果筛选出１１６个含有外源片段（长度为０．３－１ｋｂ，平均０．６ｋｂ）的克隆，经鉴定外源片段均为ＩＢＤＶｃＤＮＡ。The double stranded RNA of infectious bursal disease virus (IBDV)Qindao strain was re-verse transcribed with random primer and its cDNA libray was constructed,IBDV from thebursa of chicken was purified with immunocomplesx precipitation reaction and the dsRNA was then extracted.Following purification in agarose gel electerophoresis,the dsRNA was denaturedand reverse transcribed into cDNA.After the synthesized cDNA was tailed with dCTP,the nealed into plasmid pT7/3α-19 which was digested with Pst I and tailed with dGTP,theredcombinant plasmid was transformed into Escherichia coli strain DH5α.Based on the sereening with restriction enzyme and identification with hybridization.116 clones containingIBDV cDNA fragment (the size is 0.3-1kb,in average 0.6kb)were harvested.

关 键 词：传染性 IBD 病毒 CDNA文库 

分 类 号：S852.65[农业科学—基础兽医学]