构建由胸腺嘧啶激酶基因—小Mu噬菌体DNA插入到HSV BamHI C片段的重组质粒

为了使单纯疱疹病毒1型(HSV-1)的BamHI C片段发生突变,利用DNA重组技术将胸腺嘧啶激酶基因-小Mu噬菌体DNA系统(TK—mM)随机插入到质粒pRB 132中的HSV-1 BamHI C片段内,用DNA斑点杂交法得到6个在BamHI C片段上有TK-mM插入的重组质粒。限制性核酸内切酶的分析结果表明,由于TK—mM的插入,BamHI C片段发生突变,而TK—mM在6个重组质粒中的插入位点也是各不相同的。

CONSTRUCTION OF THE RECOMBINANT PLASMIDS CARYING BamHI C FRAGMENT OF HERPES SIMPLEX VIRUS I AND INSERTED THYMIDIN KINASE GENE-MINI Mu PHAGE DNA

In order to mutate the BamHI C fragment of Herpes Simplex Virus 1 (HSV-1), the thymidin kinase gent-mini Mu phage DNA (TK-mM) was randomly inserted into the BamHI C fragment in the plasmid pRB 132 with DNA recombination technique. The six recombinant plasmids carrying the BamHI C fragment of HSV-1 and inserted TK-mM were isolated with DNA hybridization. The results of restriction endonucleasc analysis showed that the BamHI C fragment has been mutated by insertion of TK-mM system, and the insertional sites of TK-mM located in the six recombinant plasmids were different.