中国棉铃虫核型多角体病毒VHA273毒株的克隆及基因组物理图谱

中国棉铃虫核型多角体病毒VHA273毒株的克隆及基因组物理图谱

杨忠; 黎路林; 洪华珠; 陈曲侯

华中师范大学昆虫学研究所,武汉　430070

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1996-11-08 发布日期:1996-11-08

PHYSICAL MAP OF THE GENOME OF A NUCLEAR POLYHEDROSIS VIRUS CLONE:HaNPV-H9 INFECTING HELICOVERPA ARMIGERA

YANG Zhong; LI Lu-lin; HONG Hua-zhu; CHEN Qu-hou

Institute of Entomology,Centra China Normal University,Wuhan 430070

Received:1900-01-01 Revised:1900-01-01 Online:1996-11-08 Published:1996-11-08

摘要/Abstract

摘要： 利用ＨＺ－ＩＢ３细胞对中国棉铃虫核型多角体病毒（ＨａＮＰＶ）ＶＨＡ２７３毒株进行了三轮空斑纯化，得到一个克隆株Ｈ９。电镜观察表明，此克隆株Ｈ９为单粒包埋核型多角体病毒（ＳＮＰＶ）。Ｈ９的基因组ＤＮＡ经限制性内切酶ＢｇｌＩ，ＨｉｎｄⅢ，ＰｓｔＩ，ＸｈｏＩ，ＥｃｏＲＩ，ＢａｍＨＩ，ＫｐｎＩ，ＢｓｔＥⅡ完全酶切后分别产生１０，１３，７，６，２７，１１，６和１２条片段，以λＤＮＡ／ＥｃｏＲＩ－ＨｉｎｄⅢ双酶切片段和ＨｉｇｈＭｏｌｅｃｕｌａｒＷｅｉｇｈｔＤＮＡＭａｒｋｅｒｓ为标准作工作曲线，求出了各限制性内切酶片段的大小。实验测得ＨａＮＰＶ－Ｈ９基因组的大小为１２９．９６ｋｂ，分子量为８５．７７×１０^６，表明Ｈ９不同于其它已报道的ＨａＳＮＰＶ。利用单、双酶混合酶切分析和部分酶切分析，通过逻辑推导，完成了Ｈ９基因组ＤＮＡ的ＰｓｔＩ和ＸｈｏＩ各单酶解片段的重叠和延伸，最终构建了Ｈ９基因组ＤＮＡ的ＰｓｔＩ、ＸｈｏＩ的物理图谱。

Abstract: A single-nucleiocapsid nuclear polyhedrosis virus (SNPV ) clone, designated
HaNPV-H9, has been isolated from Helicoverpa armigera NPV VHA 273 by
plaque-purification and the viral DNA of this clone was analyzed with restriction endonucleases EcoR I, BamHI, Kpn I, BstE II, BglI, Hind III, Pst I and Xho I. The sizes of the restriction fragments were calculated with respect to mobilities of standard λDNA/EoR I-Hind III f ragments and High Molecular Weight DNA Markers. The total size of the genome was about 129. 96Kb. And the restriction pattern of H9-DNA is different from the other two HaSN PV variants reported previously. The physical map of the HaN PV-H9 genome was construsted utilizating the above enzymes by multiple restriction endonucleases digestion and partialenzyme digestion. The general locations of Pst I, Xho I fragments related to each other have been determined.

中图分类号:

Q523.3

杨忠; 黎路林; 洪华珠; 陈曲侯. 中国棉铃虫核型多角体病毒VHA273毒株的克隆及基因组物理图谱[J]. , DOI: .

YANG Zhong; LI Lu-lin; HONG Hua-zhu; CHEN Qu-hou. PHYSICAL MAP OF THE GENOME OF A NUCLEAR POLYHEDROSIS VIRUS CLONE:HaNPV-H9 INFECTING HELICOVERPA ARMIGERA[J]. , DOI: .