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[甜菜夜蛾核型多角体病毒在甜菜夜蛾细胞中持续感染的建立]

[Establishment of SeMNPV persistent infection in Spotoptera exigua cells].

作者信息

Weng Qing-Bei, Xiao Wei, Yuan Mei-Jin, Yang Kai, Pang Yi

机构信息

State Key Laboratory of Biocontrol and Institute of Entomology, Sun Yat-sen University, Guangzhou 510275, China.

出版信息

Bing Du Xue Bao. 2011 Jul;27(4):347-52.

Abstract

Persistent baculovirus infection is observed frequently in insect populations. Persistent infection can be transformed to a replicative and infective state caused by stress factors and plays an important role in regulating the size of insect population and in epizoology of baculoviruses. The aim of this study is to establish a persistently baculovirus-infected cell system to explore the molecular mechanisms of baculoviral persistence. Spodoptera exigua nucleopolyhedrovirus (SeMNPV) was serially undiluted passaged in Se301 cells to reduce virulence. Upon infection of Se301 cells with the SeMNPV up to passage 8, a few cells survived even if most of cells died due to virus infection. The surviving cells were passaged and designated as P8-Se301 cell strain. P8-Se301 cells had a population doubling time of 58-65 hours and grew slower than Se301 cells. Light microscopy and electron microscopy observation showed symptom of baculovirus infection, such as virogenic stroma, viral particles and occlusion bodies, in some of P8-Se301 cells. End-point dilution assay and infectious center assay showed that 4.14% +/- 0.99% cells continually released infectious progeny virus which replicated slower than SeMNPV in Se301 cells. The result indicated that P8-Se301 cells show a typical character trait of baculovirus persistent infection.

摘要

在昆虫种群中经常观察到杆状病毒的持续感染。持续感染可因应激因素转变为复制性和感染性状态,在调节昆虫种群数量以及杆状病毒的流行病学方面发挥重要作用。本研究的目的是建立一个杆状病毒持续感染的细胞系统,以探索杆状病毒持续感染的分子机制。将甜菜夜蛾核型多角体病毒(SeMNPV)在Se301细胞中连续传代且不稀释以降低毒力。用传至第8代的SeMNPV感染Se301细胞时,即使大多数细胞因病毒感染而死亡,仍有少数细胞存活。将存活的细胞传代并命名为P8 - Se301细胞株。P8 - Se301细胞的群体倍增时间为58 - 65小时,生长速度比Se301细胞慢。光学显微镜和电子显微镜观察显示,部分P8 - Se301细胞出现杆状病毒感染症状,如病毒发生基质、病毒粒子和多角体。终点稀释试验和感染中心试验表明,4.14%±0.99%的细胞持续释放感染性子代病毒,其在Se301细胞中的复制速度比SeMNPV慢。结果表明,P8 - Se301细胞表现出杆状病毒持续感染的典型特征。

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