形成多角体家蚕重组病毒通用转移载体构建

Construction of Universal Transfer Vectors with Polyhedra Inclusion Bodies for Bombyx mori Nuclear Polyhedrosis Virus (BmNPV)

从家蚕核型多角体病毒（ＢｍＮＰＶ）通用转称载体ｐＢＫ２８３和粉纹夜蛾核型多角体病毒（ＴｎＮＰＶ）转移载体ｐＳＸＩＶＶＩ＋Ｘ３系列出发，构建了一个７．２ｋｂ能形成多角体且可以用于克隆含不同读码框外源基因的ＢｍＮＰＶ通用转移载体系列ｐＢＭＸ３．ｐＢＭＸ３系列以ＢｍＮＰＶ多角体结构基因上游的１．９ｋｂ和下游１．３ｋｂ的片段作为与ＢｍＮＰＶ基因组进行体内同源重组的同源序列；ｐＳＸＩＶＶＩ＋Ｘ３系列的ＳＸＩＶ双启动子用于外源基因的表达；ＡｃＮＰＶ的多角体基因作为重组病毒形成多角体的基因．以ＢｍＮＰＶ－ＬａｃＺ（ｏｃｃ－ｇａｌ＋）为出发株病毒。

For the conveninence of recombinant BmNPV to infect the larvae of silkworm, Bombyx mori, a series of 7.2 kb occlusion body positive (occ +)universal for foreign genes insertion in all reading frames, have been constructed based on BmNPV transfer vector pBK283 and TnNPV transfer plasmids pSXIVVI +X 3 series. The upstream homologous region for BmNPV genome recombination is 1.9 kb and the downstream region is 1.3 kb. The foreign genes are promoted by SXIV double promoters derived from SXIVVI +X 3 series and AcNPV polyhedrin gene serves for the occlusion bodies formation in recombinant viruses. Co transfected with BmNPV LacZ(occ +gal -)strain, the recombinant viruses are easily selected by 2 genetic markers:occlusion body (occ +)and white plaques(gal -).