异源多角体蛋白对家蚕核型多角体病毒粒子的包装

利用PCR方法从AcMNPV基因组DNA中分离出多角体蛋白基因 ,将该扩增片段克隆到转移载体pBacPAK8中 ,得到重组转移载体pOAc。将该质粒DNA与线性化的Bm BacPAK6病毒基因组DNA共传染BmN细胞 ,得到了能形成多角体且不产生蓝色空斑的重组病毒hp BmNPV。纯化该重组病毒的多角体颗粒 ,并对多角体蛋白、病毒核酸及多角体病毒颗粒进行分析 ,发现AcMNPV的多角体蛋白能在家蚕细胞中大量表达且能在细胞内识别家蚕核型多角体病毒并组装成多角体颗粒 ;病毒基因组DNA因部分交换 ,其酶切行为发生了相应的变化 ;电镜观察发现经AcMNPV多角体蛋白包装的家蚕核型多角体病毒的多角体颗粒大小为1 2 μm～ 2 9μm ,明显小于野生型家蚕核型多角体病毒的多角体颗粒

Package of Bombyx mori Nuclear Polyhedrosis Virus with Heterogoneous Polyhedring Protein

Cloning the segment of polyhedrin gene which was separated from AcMNPV genome DNA by PCR method into transfer vector pBacPAK8, we got recombinant transfer vector pOAc, then contransfecting BmN cells with the vector and linear virus Bm-BacPAK6, harvested recombinant virus hp-BmNPV which can produce polyhedrin but no blue plaques. Analysing of recombinant virus from polyhedrin, recombinant viral DNA and polyhedra, we confirmed that polyhedring of AcMNPV can be not only high-level expressed in BmN cells, But can recognize recombinant BmNPV DNA and assemble them to polyhedra. We observed that the pattern of restriction enzyme changed due to DNA recombination, the size of recombinant viral polyhedron (1.2-2.9 microns) was obviously smaller than those of wild-type BmNPV by electron microscopy.