Vlak J M, Schouten A, Usmany M, Belsham G J, Klinge-Roode E C, Maule A J, Van Lent J W, Zuidema D
Department of Virology, Agricultural University Wageningen, The Netherlands.
Virology. 1990 Nov;179(1):312-20. doi: 10.1016/0042-6822(90)90299-7.
A new baculovirus expression vector based upon the p10 gene of Autographa californica nuclear polyhedrosis virus (AcNPV) and a novel system for the screening of p10 recombinants have been developed. The insertion of a cassette containing the lacZ gene under the control of a heat-shock promoter of Drosophila melanogaster downstream from the cloning site in p10 transfer vectors allows the convenient identification of putative recombinants by virtue of their expression of beta-galactosidase. Using this p10 transfer vector an AcNPV recombinant was engineered with a cDNA copy of gene I of cauliflower mosaic virus (CaMV) in place of the p10 coding sequence. This p10 recombinant expressed CaMV gene I at levels equivalent to those of p10 and polyhedrin, and was shown to be as effective in producing this protein as recombinants exploiting the polyhedrin promoter. CaMV gene I protein formed large numbers of hollow fiber-like structures in the cytoplasm of infected cells. Because the polyhedrin gene remains intact, these p10 expression vectors may be exploited for the expression of heterologous proteins in insects infected per os and for the enhancement of baculovirus pathogenicity for insect control.
基于苜蓿银纹夜蛾核型多角体病毒（AcNPV）的p10基因构建了一种新型杆状病毒表达载体，并开发了一种筛选p10重组体的新系统。在p10转移载体的克隆位点下游插入一个含有受黑腹果蝇热休克启动子控制的lacZ基因的盒式结构，可凭借β-半乳糖苷酶的表达方便地鉴定推定的重组体。利用这种p10转移载体构建了一种AcNPV重组体，用花椰菜花叶病毒（CaMV）基因I的cDNA拷贝取代了p10编码序列。这种p10重组体表达CaMV基因I的水平与p10和多角体蛋白相当，并且在产生该蛋白方面与利用多角体蛋白启动子的重组体一样有效。CaMV基因I蛋白在受感染细胞的细胞质中形成大量中空纤维状结构。由于多角体蛋白基因保持完整，这些p10表达载体可用于在经口感染的昆虫中表达异源蛋白，并增强杆状病毒对昆虫控制的致病性。
