Zhao Ying, Yu Huan, He Lei, Li Ni, Huang Guo-Hua
Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Changsha, 410128, China.
College of Plant Protection, Hunan Agricultural University, Changsha, 410128, China.
Virus Genes. 2019 Oct;55(5):688-695. doi: 10.1007/s11262-019-01679-7. Epub 2019 Jun 24.
The open reading frame 117 (3h-117) of Heliothis virescens ascovirus 3h (HvAV-3h), which is a conserved coding region present in all completely sequenced ascovirus members, was characterized in this study. By RT-PCR detection, 3h-117 transcription began at 6-h post-infection (hpi) and remained stable until 168 hpi in HvAV-3h-infected Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae. In addition, 3h-117 putatively encodes a 21.5-kDa protein (3H-117) predicted to be a CTD-like phosphatase. Western blot analysis using a prepared rabbit polyclonal antibody specific to 3H-117 showed that the product could be detected at 24 hpi, which remained stably detectable until 168 hpi. The same analysis also demonstrated that the 3H-117 protein localized in the virions of HvAV-3h. Immunofluorescence analysis showed that at 24 hpi, 3H-117 was mainly located in the nuclei of H. armigera larval fat body cells and later spread into the cytoplasm. In summary, our results indicate that 3H-117 is a structural protein of HvAV-3h.
烟芽夜蛾浓核病毒3h（HvAV-3h）的开放阅读框117（3h-117）是所有已完成全序列测定的浓核病毒成员中存在的一个保守编码区，本研究对其进行了特性分析。通过逆转录聚合酶链反应（RT-PCR）检测，在受HvAV-3h感染的棉铃虫（鳞翅目：夜蛾科）幼虫中，3h-117转录在感染后6小时（hpi）开始，并一直稳定到168 hpi。此外，3h-117推测编码一种预测为CTD样磷酸酶的21.5 kDa蛋白（3H-117）。使用制备的针对3H-117的兔多克隆抗体进行的蛋白质免疫印迹分析表明，该产物在24 hpi时可被检测到，并且一直稳定可检测到168 hpi。同样的分析还表明，3H-117蛋白定位于HvAV-3h的病毒粒子中。免疫荧光分析表明，在24 hpi时，3H-117主要位于棉铃虫幼虫脂肪体细胞的细胞核中，随后扩散到细胞质中。总之，我们的结果表明3H-117是HvAV-3h的一种结构蛋白。
