ACTIVATION AND DETECTION OF A LATENT BACULOVIRUS RESEMBLING MAMESTRA-BRASSICAE NUCLEAR POLYHEDROSIS-VIRUS IN M-BRASSICAE INSECTS
ACTIVATION AND DETECTION OF A LATENT BACULOVIRUS RESEMBLING MAMESTRA-BRASSICAE NUCLEAR POLYHEDROSIS-VIRUS IN M-BRASSICAE INSECTS
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DOI：
10.1006/viro.1993.1300
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发表时间：
1993-06-01
期刊：
影响因子：
3.7
通讯作者：
KING, LA
中科院分区：
文献类型：
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作者：
HUGHES, DS;POSSEE, RD;KING, LA
实验室培养的甘蓝夜蛾（Mamestra brassicaeinsects，MbLC）被发现潜伏杆状病毒感染。用M.甘蓝型油菜夜蛾与近缘的Panolis火焰核型多角体病毒（NPV）或远缘的Autographa californica NPV的关系。经酶切分析表明，该病毒与M病毒的亲缘关系很近，甚至不完全相同。甘蓝核型多角体病毒多角体蛋白基因序列的聚合酶链反应扩增表明，潜伏病毒存在于整个生命周期的昆虫，卵，幼虫，蛹，和成人。我们未能在第二次培养的M.从环境中获得并且最近才适应在实验室条件下生长的芸苔属昆虫，因此这些昆虫在所有PCR和活化实验中充当有效的阴性对照。用PCR分析从四龄MbLC幼虫解剖组织中分离的DNA，仅在脂肪体中检测到潜伏病毒序列。从分离的MbLC脂肪体组织中建立的细胞系也显示出潜伏病毒序列，并且应该证明在进一步的研究中是有用的，以阐明潜伏和病毒活化的机制。
A laboratory culture ofMamestra brassicaeinsects (MbLC) has been found to harbor a latent baculovirus infection. The latent virus was activated by feeding theM. brassicaelarvae with either the closely relatedPanolis flammeanuclear polyhedrosis virus (NPV), or the distantly relatedAutographa californicaNPV. Restriction fragment profiles of the activated virus DNA showed that it is very closely related, if not identical, toM. brassicaeNPV. Polymerase chain reaction amplification of polyhedrin gene sequences demonstrated that the latent virus was present throughout the life cycle of the insect; eggs, larvae, pupae, and adults. We failed to detect the presence of a latent virus in a second culture ofM. brassicaeinsects, obtained from the environment and only recently adapted to growth in laboratory conditions, and thus these insects acted as an effective negative control in all the PCR and activation experiments. Using PCR analysis of DNA isolated from dissected tissues of fourth instar MbLC larvae, latent virus sequences were only detected in the fat body. Cell lines established from the isolated MbLC fat body tissue were also shown to harbor the latent virus sequences and should prove useful in further studies to elucidate the mechanisms of latency and virus activation.