PCR介导HaNPV罹病幼虫潜伏期cDNA文库的构建  

Construction of Helicoverpa armigera Nuclear Polyhedrovirus cDNA Library During Latent Periods by Polymerase Chain Reaction

作　　者：白志强[1] 张友清[1] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071

出　　处：《中国病毒学》2000年第4期346-349,共4页Virologica Sinica

摘　　要：以罹病棉铃虫幼虫为材料提取总RNA ,反转录合成cDNA第一链 ,加oligo(dG)同聚尾 ,PCR扩增合成双链cDNA ,克隆到 pGEM T质粒载体中。随机筛选文库中阳性克隆 ,经酶切分析 ,cDNA插入片段大小在 0 .3～ 1.1kb之间。文库中原代重组子数为 1.66× 10 5,重组百分比为 87.8%。重组质粒的杂交分析表明 ,文库中HaNPV基因的cDNA克隆所占比例超过 50 %。A general cDNA library which was from infected larvae dueing the latent periods has been constructed. The first cDNA strand was synthesized from total RNA by M MLV transcriptase. After oligo(dG) tailing the cDNA was amplified by polymerase chain reaction. The double strands cDNA were ligated to pGEM T easy vectors. Screeing of cDNA library showed that the length of inserts was from 0.3 to 1.1 kilobases. The capacity of cDNA library was 1.66×10 5 clones. Moreover, the number of cDNA clones of HaNPV was more than 50%.

关 键 词：中国棉铃虫 核型多角体病毒 克隆 CDNA文库 

分 类 号：Q78[生物学—分子生物学]