稳定表达绿色荧光蛋白的盖塔病毒的构建与鉴定  

Reconstruction and characterization of a stable Getah virus strain expressing green fluorescent protein

作　　者：姜智文 秦颖 陈杰[1] 粟硕[1] JIANG Zhiwen;QIN Ying;CHEN Jie;SU Shuo(Sanya Institute of Nanjing Agricultural University,Sanya 572000,China)

机构地区：[1]南京农业大学三亚研究院,海南三亚572000

出　　处：《畜牧与兽医》2025年第3期74-78,共5页Animal Husbandry & Veterinary Medicine

基　　金：三亚崖州湾科技城科技专项资助项目(SCKJ-JYRC-2022-08)。

摘　　要：为了开发表达外源基因的重组盖塔病毒,通过反向遗传技术,在盖塔病毒nsP3基因的C端插入绿色荧光蛋白ZsGreen,并通过转染至BHK-21细胞拯救重组病毒并研究其生长特性。结果:感染性cDNA克隆pSM-GETV-nsP3/ZsGreen转染BHK-21细胞后表达绿色荧光蛋白,所拯救的报告病毒可以在BHK-21细胞上稳定传代(至少10代),并表现出良好的遗传稳定性;多步生长曲线试验表明,报告病毒GETV-ZsGreen在BHK-21细胞上的增殖能力略低于亲本毒株,但仍能达到较高病毒滴度。综上,本研究构建并拯救了1株遗传稳定的GETV报告病毒GETV-ZsGreen。In order to develop a recombinant Getah virus capable of expressing exogenous genes,this study employed reverse genetic technology to insert the ZsGreen protein at the C-terminus of the nsP3 gene of Getah virus by transfecting a cDNA clone pSM-GETV-nsP3/Zs-Green into BHK-21 cells and investigated its growth characteristics.Our findings demonstrated that,upon transfection into BHK-21 cells,the infectious cDNA clone pSM-GETV-nsP3/ZsGreen expressed green fluorescent protein,and subsequently rescued reporter virus exhibiting stable propagation(for at least 10 passages)in BHK-21 cells with excellent genetic stability.The multi-step growth curve analysis revealed that GETV-ZsGreen reporter virus displayed slightly reduced replication ability compared with its parental strain in BHK-21 cells;nevertheless,it still achieved a high viral titer.In summary,we successfully constructed and rescued a genetically stable GETV report virus,designated GETV-ZsGreen.

关 键 词：盖塔病毒 感染性克隆 报告病毒 

分 类 号：S852[农业科学—基础兽医学]