柞蚕核多角体病毒核多角体基因的定位和克隆

通过Southern转印杂交证明,柞蚕核多角体病毒(Antheraea pernyi nuclear polyhedrosis virus,ApNPV)核多角体基因位于该病毒基因组DNA Bam HⅠ D和E片段上,我们巳将这两个片段分别克隆到pAT153质粒中,并用末端杂交法确定了ApNPV核多角体基因的方向,对含有这一基因的片段进行了限制性内切酶图谱分析,进而对这一基因部分编码区进行了核苷酸序列分析,在用ApNPV这一段序列(222bp)与其他昆虫核多角体病毒AcNPV(AutograPha californica NPV,苜蓿丫纹夜蛾NPV);BmNPV(Bombyx mory NPV,家蚕NPV);OpNPV(Orqyia Pseudotsugata NPV,黄杉毒蛾NPV)核多角体基因相应区段相比较分析中,发现它们之间的同源核苷酸序列比率分别为77.5％、84％和80％。

LOCATION AND CLONING OF POLYHEDRIN GENE OF ANTHERAEA PERNYI NUCLEAR POLYHEDROSIS VIRUS

The polyhedrin gene wasi dentified to be located on the BamH I D and E fragments of ApNPV genome DNA by Southern hybridization with OpNPV polyhedrin gene fragment as a probe. Subsequently, the BamH I D and E fragments were cloned into plasmid AT153. A portion containing the polyhedrin gene of ApNPV was mapped and the 5' and 3'ends of the gene were determined by the terminal hybridization procedure. Furthermore,a small region containing 222 bp coding for ApNPy polyhedrin gene was sequenced.The result indicated that the nucleotide sequences of ApNPV polyhedrin gene is homologous with AcNPV, BmNPV and OpNPV by 75.5%,84% and 80% repectively.