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野生型甜菜夜蛾核多角体病毒 (SeMNPV)US1分离株 (SeUS1 )通过空斑法纯化 ,PCR、长片段PCR和限制性内切酶分析筛选和鉴定 ,获得一株基因型较为均一且具完整基因组的克隆株 ,命名为Se 4。Se 4在其宿主细胞系Se3 0 1中无稀释连续传代至 1 0代 ,各代被感染细胞中的病毒DNA经限制性内切酶分析 ,发现在第 7代时病毒基因组中出现了一条新增的 3 .5kb片段 ,随着代数的增加 ,该片段的摩尔量逐渐增加 ,在第 1 0代时已成为超摩尔带 ,推测该片段为SeMNPVDNA复制的顺式作用元件。序列分析表明 ,该片段覆盖了SeMNPV 81 0 1 4～ 845 3 8nt共 3 5 2 5bp的序列 ,包含被预测为杆状病毒的DNA复制原点的non hr区域以及一些SeMNPV特有的ORF。研究结果为核多角体病毒II组的non hr在病毒复制过程中具有重要作用的观点提供了体外实验的证据
The wild-type S. litura nuclear polyhedrosis virus (SeMNPV) US1 isolate (SeUS1) was purified by plaque assay, and screened and identified by PCR, long PCR and restriction endonuclease analysis to obtain a more homogenous and complete genotype The cloned strain of the genome, designated Se 4. Se 4 was continuously passaged to 10 generations without dilution in its host cell line Se3 0 1. The viral DNA in the infected cells of each passage was analyzed by restriction enzyme analysis and found a new gene in the virus genome at the 7th generation With the increase of algebra, the molar amount of this fragment increased gradually and became a supermolecular band at the10th generation. It was presumed that this fragment was the cis-acting element of SeMNPVDNA replication. Sequence analysis showed that the fragment covered a sequence of SeMNPV 81 0 1 4 ~ 845 3 8nt with 3 5 2 5 bp, including the non hr region predicted to be the origin of baculovirus DNA replication and some SeMNPV-specific ORFs. The results provide evidence of in vitro experiments for the view that non hr nuclear polyhedrosis virus group II has an important role in viral replication