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犬Ⅰ型腺病毒DNA的酶切分析及分子克隆 被引量:2

Restriction Enzyme Analysis and Molecular Cloning of CAV-1 DNA
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摘要 犬Ⅰ型腺病毒(CAV-1)弱毒用限制性内切酶EcoR Ⅰ,BamH Ⅰ,Pst Ⅰ,Sph Ⅰ和Hind Ⅲ消化分析后其图谱与强毒株相比没有差异。将弱毒DNA用Pst Ⅰ完全消化后以鸟枪法克隆到载体质粒pBluescrip'SK中,经用光生物素标记的CAV-1 DNA杂交筛选以及Pst Ⅰ分析重组质粒证明已将分子量为5.5,3.5,2.85,1.2,0.32和0.28Kb的CAV-1DNA片段克隆到质粒中。克隆到的这些片段将可考虑进一步研究作为探针检测犬及狐狸等野生动物的腺病毒感染。 The virulent and modified canine adenovirus type 1 (CAV-1) DNAs extracted from canine kidney cells (MDCK) 36 hrs postinfection were both analysed with restriction enzymes EcoR Ⅰ, BamH Ⅰ, Pst Ⅰ, Nsi Ⅰ, Sph Ⅰ and Hind Ⅲ. and no difference was found from the electrophoresis pattern between the two viruses. The modified CAV-1 DNA was digested completely with Pst Ⅰ and cloned into, plasmid pBluescript SK taking shotgun procedure. Through hybridization selection of photobiotin labeled CAV-1 DNA and Pst Ⅰ digestion of recombinant plasmids, 6 CAV-1 DNA Pst Ⅰ fragments with moI. wets. of 5.5, 3.5, 2.85, 1.2, 0.32 and 0.28kb, were indicated being cloned in the plasmid. These clones will be used as probes to diagnose CAV-1 infections for dogs and foxes.
出处 《中国病毒学》 CSCD 1992年第3期357-361,共5页 Virologica Sinica
关键词 犬I型腺病毒 DNA酶切分析 克隆 CAV-1 DNA fingerprinting Molecular cloning
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