牛疱疹病毒Ⅳ型(DN-599株)DNA的限制性酶切位点图及重复序列分析

牛疱疹病毒Ⅳ型(BHV-4)DNA片段分别被重组到质粒pUC9或pBR322的EcoRI、Hind Ⅲ和BamHⅠ位点中,用光生物素(Photobiotin)标记这些重组质粒作为探针,分别与转移到硝基纤维素膜上的病毒DNA的EcoRⅠ、Hind Ⅲ和BamHⅠ片段进行杂交,根据杂交结果及克隆的BHV-4DNA片段的双酶切分析,画出了病毒DNA的EcoRⅠ、Hind Ⅲ和BamHⅠ位点图,井证明在病毒DNA的两末端含有多聚重复序列,左侧含12个重复单位,右侧含6个重复单位,两侧重复单位的排列方向相同,重复单位的大小为2.35kb。病毒DNA分子无任何异构体.

PHYSICAL MAPPING OF BOVINE HERPESVIRUS TYPE 4 ( DN-599 STRAIN ) DNA AND ANALYSIS OF ITS REPETITIVE SEQUENCES

Bovine herpesvirus type 4 ( BHV-4)DNA has been cloned into EcoR I, Hind III, or BamH I sites of pUC9 or pBR322 respectively. The recombin-ant plasmids harboring BHV-4 DNA fragments labeled with photobiotin were used as probes to hybridize to each of EcoR I, Hind I and BamH I digests of BHV-4 DNA on the nitrocellulose membrane which had been transferred from agarose gel after electrophoresis. With the hybridization and double digesticn of cloned BHV-4 DXA fragments, three physical maps of BHV-4 DNA of EcoR I, Hind I and BamH I are presented. There are po-lyrepetitive sequences at both termini of the viral genome in the same orientation, 12 repeated units on the left end and 6 on the right end. The size of repeat unit is 2.35kb. The viral DNA has no isomers.