Molecular Cloning of Bovine Herpesvirus Type 4 (DN-599 Strain) DNA

TONG Guang-Zhi; Dieter Burger; David T Shen

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August 1990

TONG Guang-Zhi, Dieter Burger and David T Shen. Molecular Cloning of Bovine Herpesvirus Type 4 (DN-599 Strain) DNA[J]. Virologica Sinica, 1990, 5(4).

Citation: TONG Guang-Zhi, Dieter Burger, David T Shen. Molecular Cloning of Bovine Herpesvirus Type 4 (DN-599 Strain) DNA .VIROLOGICA SINICA, 1990, 5(4) : 423.

牛疱疹病毒Ⅳ型(DN-599株)DNA的分子克隆

1.
中国农业科学院哈尔滨兽医研究所
2.
美国华盛顿州立大学USDA-ARS 实验室
3.
美国华盛顿州立大学USDA-ARS 实验室哈尔滨

摘要

从感染的MDBK(牛肾)细胞中直接抽提出牛疱疹病毒IV型(BHV-4)DNA。分别经限制性内切酶EcoRⅠ,HindⅢ或BamHI消化后,用鸟枪法和选择法将病毒DNA片段克隆到质粒pBR322和pUC9的相应位点中,构建了三组病毒DNA基因库。阳性克隆是用光生物素标记的病毒DNA和牛胸腺细胞DNA与各重组质粒DNA杂交而筛选的。总共克隆了病毒基因组的94％,其中用EcoRI克隆了83.4％,BamHI克隆了63.4％,HindIlI克隆了45％
- 牛疱疹病毒IV型
- , 分子克隆
- , 光生物素

Molecular Cloning of Bovine Herpesvirus Type 4 (DN-599 Strain) DNA

1.
Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences
2.
Harbin 150001USDA-ARS
3.
Washington State University

Abstract

Bovine herpesvirus type 4 (BHV-4) DNA was extracted from infected MDBK (Bovine kidney) cells using Triton Ⅹ-100 and phenol/chloroform/ether. With shotgun and selective cloning procedures, the viral DNA was cloned into EcoR I, BamH I or Hind III sites of plasmids pBR322 or pUC9 after digestion of the DNA with the three restriction endonucleases, respectively, and three DNA libraries containing different restricted fragments were constructed. Photobiotin-labeled viral DNA and bovine thymus DNA were used as pr...
- BHV-4
- , Molecular cloning
- , Photobiotin

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Molecular Cloning of Bovine Herpesvirus Type 4 (DN-599 Strain) DNA

1. Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences
2. Harbin 150001USDA-ARS
3. Washington State University

Abstract: Bovine herpesvirus type 4 (BHV-4) DNA was extracted from infected MDBK (Bovine kidney) cells using Triton Ⅹ-100 and phenol/chloroform/ether. With shotgun and selective cloning procedures, the viral DNA was cloned into EcoR I, BamH I or Hind III sites of plasmids pBR322 or pUC9 after digestion of the DNA with the three restriction endonucleases, respectively, and three DNA libraries containing different restricted fragments were constructed. Photobiotin-labeled viral DNA and bovine thymus DNA were used as pr...