Gupta P K, Chaudhuri P, Sharma B, Singh V P, Balain D S
National Biotechnology Centre Indian Veterinary Research Institute, UP, India.
Biochem Mol Biol Int. 1995 Jan;35(1):167-75.
The DNA from an Indian isolate of Bovine Herpesvirus-1 was isolated and analysed with restriction endonucleases. On shot gun cloning seven HindIII digested BHV-1 DNA fragments could be cloned in pBR322 vector. Recombinant clones with viral DNA insert was identified by insertional inactivation of the selection marker and restriction analysis of recombinant plasmids with HindIII. Further, recombinant plasmids were analysed with HindIII, EcoRI and BamHI restriction endonucleases to identify the different viral DNA inserts. The restriction site map of recombinant plasmids were correlated with the map reported earlier. The southern blot of restriction digested recombinant plasmids was hybridized with radio-labelled BHV-1 DNA as probe.
从一株印度牛疱疹病毒1型(BHV-1)分离物中提取DNA,并使用限制性内切酶进行分析。通过鸟枪法克隆,七个经HindIII酶切的BHV-1 DNA片段可克隆到pBR322载体中。通过选择标记的插入失活以及用HindIII对重组质粒进行限制性分析,鉴定出带有病毒DNA插入片段的重组克隆。此外,用HindIII、EcoRI和BamHI限制性内切酶对重组质粒进行分析,以鉴定不同的病毒DNA插入片段。将重组质粒的限制性酶切位点图谱与先前报道的图谱进行比对。用放射性标记的BHV-1 DNA作为探针,对经限制性酶切的重组质粒进行Southern杂交。