ISOLATION OF GENOTYPIC VARIANTS OF AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS
ISOLATION OF GENOTYPIC VARIANTS OF AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS
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DOI：
10.1128/jvi.27.3.754-767.1978
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发表时间：
1978-01-01
影响因子：
5.4
通讯作者：
MILLER, LK
中科院分区：
文献类型：
--
作者：
LEE, HH;MILLER, LK
以只含1个核衣壳/被膜的病毒粒子为接种物，通过连续空斑纯化，从鳞翅目昆虫中分离到一株核型多角体病毒(MNPV)。通过分离具有相似但不完全相同的限制性内切酶片段的病毒的无缺陷的基因变异体，证明了用该方法克隆病毒的能力。用HindIII、EcoRI、Sali和BamHI限制性内切酶进行基因分型，鉴定出5个不同的变异体。在昆虫幼虫中传代后，每个变异体的特征基因都保持不变。这些病毒变异体的分离表明了未克隆病毒制剂的异质性，以及通过对介质来源的非包涵体病毒粒子进行空斑纯化来克隆MNPV的能力。美国正考虑将斑潜蝇MNPV作为一种杀虫剂用于商业用途，鉴于检测到的异质性，克隆该病毒可能是可取的。克隆和基因分型分析对于理解多重包埋核型多角体病毒(包膜形式中含有超过1个核衣壳/包膜的核型核型多角体病毒)的遗传性质也具有重要意义，并表明对这些病毒进行进一步的遗传分析是可能的。
A nuclear polyhedrosis virus (MNPV) isolated from a lepidopteran (Noctuidae) insect, A. californica, was cloned by successive plaque purification using virions containing only 1 nucleocapsid/envelope as inoculum. The ability to clone the virus by this method was demonstrated by the isolation of nondefective, genotypic variants of the virus with similar but not identical restriction endonuclease fragment patterns. Five distinct variants were identified by genotypic analysis with HindIII, EcoRI, SalI and BamHI restriction endonucleases. The characteristic genotype of each variant was maintained upon passage in insect larvae. The isolation of these virus variants demonstrates the heterogeneity of the uncloned virus preparation and the ability to clone MNPV by plaque purification of media-derived nonoccluded virions. The A. californica MNPV is being considered for commercial use as a pesticide in the USA, and the cloning of the virus, in view of the heterogeneity detected, may be advisable. The cloning and genotypic analyses are also significant with regard to understanding the genetic nature of multiply embedded NPV (those NPV containing more than 1 nucleocapsid/envelope in the occluded form of the virus) and indicate that further genetic analysis of these viruses is possible.