斜纹夜蛾NPV多角体基因的克隆和部分测序  

Clone and Partial Nucleotide Sequence of the Spodoptera litura Nuclear Polyhedrosis Virus Polyhedrin Gene

作　　者：曾庆[1] 龙綮新[1] 王章[1] 蒲蛰龙[1] 

机构地区：[1]中山大学昆虫学研究所/生物防治国家重点实验室,广东省微生物研究所

出　　处：《中山大学学报（自然科学版）》1994年第4期70-77,共8页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金资助项目

摘　　要：本文对ＳＩＮＰＶ基因组作了酶解分析，测得其基因组大小为１４５ｋｂ，并用双酶法确定了ＳＩＮＰＶ基因组的ＨｉｎｄⅢ和ＰｓｔⅠ物理图谱。以含ＡｃＮＰＶ多角体基因的质粒ｐＡＣ－Ⅰ的ＳａｌＩ－Ｃ片段为探针，对ＳＩＮＰＶＤＮＡ酶切片段ｓｏｕｔｈｅｒｎ转印杂交结果，初步判断多角体基因定位于ＰｓｔＩ－Ｂ／Ｃ／Ｄ片段、ＢｇｌⅡ－Ｃ／Ｄ片段、ＢａｍＨＩ－Ｂ／Ｃ片段和ＥｃｏＲＩ－Ａ／Ｂ片段上，且ＳＩＮＰＶ与ＡｃＮＰＶ多角体蛋白基因有６４％的同源性，而以大肠仟菌质粒ｐＵＣ１９为载体对ＳＩＮＰＶ的多角体基因试克隆，得到带有ＢｇｌⅡ－ＰｓｔⅠ双酶切片段的２个克隆子。对这两个杂交阳性克隆子之一的核苷酸序列测定，表明插入片段与ＢｍＮＰＶ多角体基因上游序列亦有一定同源性。A restriction endonuclease map was determined for the Splodopera litura nuclear polyhe-drosis virus(SINPV ) genome. The order of the fragments generated by the enzymes Hind Ⅲ andPst I was analysed using double digestion of the total genome and digested of isolated restrictionfragments. The Iocation of the polyhedrin gene was then determined using a cloned polyhedringene from the Autographa californica NPV as a hybridization probe. The SINPV polyhedrin genewas located on Pst I- B/C/D,BamH I- B/Cand EcoR I- A/B fragments. A fragrnent con-taining this gene was cloned and the pratial nucleotide sequence of a 1.okbfragment was deter-mined which contained the plyhedrin reading frame or/and some flanking sequences. This genewas demonstrated 64% nucleotide sequence homology to the AcNPV polyhedrin gene and 53.6%～59.6% nucleotide sequence homology to the Bombyx mori NPV polyhedrin gene.

关 键 词：斜纹夜蛾 NPV 多角体基因 克隆 

分 类 号：Q969.436.5[生物学—昆虫学]