斜纹夜蛾核型多角体病毒p^(74)基因的克隆和序列分析

Sequencing of the p^(74) Gene of Spodoptera litura Multinucleocapcid Polyhedrosis Virus

用AcNPV p74基因 3′端的 1.4kb片段 ,通过Southernblotting将SpltNPV的 p74基因定位于XhoI(4 .9kb)、EcoRI(4 .4kb)、BamHI(3.0kb)片段上。进一步用ExoⅢ和构建亚克隆测序法 ,对长 2 545bp的 p74基因进行了序列分析 ,其中 1974bp的编码区编码 6 58个氨基酸 ,蛋白分子量约 75.87kD ,其编码蛋白与AcMNPV和CfMNPV p74蛋白的氨基酸序列同源性分别为 6 4 %和6 3%。

The p74 gene of SpltNPV was located at EcoRI 4.4 kb,XhoI 4.9 kb and BamHI 3.0 kb fragments by Southern blotting with AcMNPV derived probe. After cloning of the three fragments, restriction enzyme analysis was used to construct their restriction map. A region of 2?545 bp including p74 gene of SpltNPV was sequenced. The open reading frame of SpltNPV p74 gene was decided to be 1?974 bp long, which codes 658 amino acids. Comparison of the homology with other baculovirus p74 genes shows that SpltNPV p74 is 64% and 63% identical to that of AcMNPV and CfMNPV at the amino acid level.