异源包装家蚕核型多角体病毒的特性研究

STUDY ON THE PROPERTIES OF BOMBYX MORI NUCLEAR POLYHEDROSIS VIRUSWITH HETEROGENEOUS POLYHEDRIN

利用PCR方法分离AcMNPV多角体蛋白基因及其启动子的特异片段 ,将该基因片段正确克隆到转移载体pBacPAK(API 4)中 ,得到重组转移载体pBacOAc(API4)。将它与已缺失多角体蛋白基因的BmNPV(即 :CrBK)基因组DNA共转染BmN细胞 ,将AcMNPV多角体蛋白基因及慈菇蛋白酶抑制剂基因定向克隆到CrBK基因组中 ,得重组病毒BmNPV(OAc -API4)。发现该重组病毒能被AcMNPV多角体蛋白所识别 ,并形成多角体包埋型病毒 ;CrBK粒子能被正确包装 ,同时也能表达慈菇蛋白酶抑制剂。比较了该重组病毒在家蚕细胞及家蚕幼虫中的增殖情况 ,发现它的芽生型病毒能正常感染家蚕细胞和经皮下注射的家蚕幼虫 ,然而其包埋型病毒不能经口感染家蚕幼虫。表明NPV的经口感染与多角体蛋白的种类有关。

The polyhedrin gene and its promoter of Autographa californica multicapsid nuclear polyhedrosis virus(AcMNPV) was separated by PCR.Then cloned it into transfer vector pBacPAK(API4) and obtained recombinant transfer vector pBacOAc(API4).Contransfected in BmN cells with CrBK genome which has been deleted the polyhedrin gene of Bombyx mori nuclear polyhedrosis virus.And a recombinant BmNPV(OAc-API4) was obtained,which was able to produce AcMNPV polyhedrin protein and expressed the Arrowhead proteinase inhibitor.The replication of the recombinant virus in silkworm cells and larvae showed that the budded virus could infect BmN cells and injected silkworm larvae,while the per os inoculation with its occluded virus couldn′t infect the larvae.It was indicated that the per os inoculation of NPV was concerned with the type of polyhedrin proteins.