烟芽夜蛾囊泡病毒3h毒株3h-161的克隆和表达谱分析  

Cloning and expression of 3h-161 gene encoded by Heliothis virescens ascovirus 3h(HvAV-3h)

作　　者：陈斌[1] 徐存翊 黎妮[1] 谭赟 于欢 黄国华[1] CHEN Bin;XU Cun-Yi;LI Ni;TAN Yun;YU Huan;HUANG Guo-Hua(Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学植物病虫害生物学与防控湖南省重点实验室,长沙410128

出　　处：《应用昆虫学报》2022年第3期599-607,共9页Chinese Journal of Applied Entomology

基　　金：国家自然科学基金项目(31872027);国家现代农业产业技术体系(CARS-23-C08)。

摘　　要：【目的】本文旨在明确烟芽夜蛾囊泡病毒3h毒株(HvAV-3h)编码的第161号开放阅读框(3h-161)的序列特征及表达谱,为深入研究其功能奠定基础。【方法】采用PCR技术克隆3h-161的CDs序列,构建原核表达载体,并制备高效的多克隆抗体;利用RT-PCR技术检测3h-161的转录情况,利用蛋白免疫印迹法检测3H-161的表达情况。【结果】3h-161编码区长516 bp,共编码172个氨基酸残基,感染甜菜夜蛾Spodopteraexigua3龄幼虫后12h开始转录、48h开始表达;3H-161在囊泡病毒属内的不同毒株(HvAV-3j、HvAV-3i、HvAV-3e、HvAV-3f和HvAV-3g)间高度保守,序列一致性为100%。【结论】本研究阐明了3h-161的序列特征、获得了3H-161的多克隆抗体,明确了其表达谱,为进一步研究该基因在HvAV-3h感染甜菜夜蛾过程中发挥的作用奠定基础。[Objectives] To clarify the sequence characteristics and expression profile of the 3h-161(open reading frame 161) gene in Heliothis virescens ascovirus 3h(HvAV-3h), thereby laying a foundation for further study of the function of this gene. [Methods] The CDs sequence of 3h-161 was cloned using PCR technology, a prokaryotic expression plasmid vector constructed and efficient polyclonal antibody prepared. RT-PCR and Western blotting technology was then used to detect the transcription and expression of 3H-161. [Results] The coding regions of 3h-161 are 516 bp in length, encoding 172 amino acids, respectively. Transcription began at 12 h and expression from 48 h in 3rd instar S. exigua larvae after infection with HvAV-3h. The putative protein sequences of 3h-161 are highly conservative in the Ascovirus and share 100% amino acid sequence identity with HvAV-3j, HvAV-3i, HvAV-3e, HvAV-3f and HvAV-3g. [Conclusion] The sequence characteristics of 3h-161 were determined, a polyclonal antibody of 3H-161 was obtained, and the expression profile of this gene was clarified,all of which lay a foundation for further study of the function of the 3h-161 gene during the infection of S. exigua by HvAV-3h.

关 键 词：烟芽夜蛾囊泡病毒3h毒株(HvAV-3h) 3h-161 原核表达 时相分析 

分 类 号：S432.41[农业科学—植物病理学]