Tomita S, Kanaya T, Kobayashi J, Imanishi S
Department of Insect Genetics and Breeding, National Institute of Sericultural and Entomological Science, Ibaraki, Japan.
Cytotechnology. 1995;17(2):65-70. doi: 10.1007/BF00749393.
A homologue of Autographa californica NPV (AcNPV) p10 gene was identified and cloned from Bombyx mori NPV (BmNPV). BmNPV p10 gene encodes truncated protein of 70 amino acid residues that lacks carboxyl terminus comparing with the p10 protein encoded by AcNPV. The putative TATA box sequence and the ATAAG motif which is the consensus sequence of baculovirus very late promoter were conserved. A transfer vector, pBNT1, which includes the p10 promoter region of BmNPV for foreign gene expression was constructed. By using pBNT1, a recombinant BmNPV, Bmp10-Luc, in which the p10 gene was replaced by the firefly luciferase gene, was obtained. We also obtained another recombinant virus, BmPH-Luc, in which the polyhedrin gene was replaced by the luciferase gene. The luciferase activity detected in BoMo-15AIIc insect cells infected with Bmp10-Luc was approximately 50% of that infected with BmPH-Luc, suggesting that although both the p10 and polyhedrin promoters of BrnNPV are effective in high-level expression of foreign genes, the p10 promoter is not so strong as the polyhedrin promoter.
从家蚕核型多角体病毒（BmNPV）中鉴定并克隆了苜蓿银纹夜蛾核型多角体病毒（AcNPV）p10基因的同源物。BmNPV p10基因编码一个由70个氨基酸残基组成的截短蛋白，与AcNPV编码的p10蛋白相比，该蛋白缺少羧基末端。推测的TATA框序列和杆状病毒晚期启动子共有序列ATAAG基序是保守的。构建了一个用于外源基因表达的转移载体pBNT1，其包含BmNPV的p10启动子区域。利用pBNT1，获得了一种重组BmNPV，即Bmp10-Luc，其中p10基因被萤火虫荧光素酶基因取代。我们还获得了另一种重组病毒BmPH-Luc，其中多角体蛋白基因被荧光素酶基因取代。在感染Bmp10-Luc的BoMo-15AIIc昆虫细胞中检测到的荧光素酶活性约为感染BmPH-Luc的细胞的50%，这表明尽管BmNPV的p10和多角体蛋白启动子在外源基因的高水平表达中均有效，但p10启动子不如多角体蛋白启动子强。
