形成多角体家蚕重组病毒通用转移载体构建

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，ｐＢＭＸ３系列的重组病毒筛选有ｏｃｃ＋和ｇａｌ－两个遗传标记

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

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 -).