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％。
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 probes to identify the recombinant plasmids which are harbouring the viral DNA fragments. Total 94% of the entire genome of BHV-4 (83.4% with EcoR I, 63.4% vvith BamH I and 45% with HindIII) was cloned.
出处
《病毒学杂志》
CSCD
1990年第4期423-429,共7页
