【精品论文】荧光素酶标记狂犬病病毒的反向遗传学.pdf

936 2010 7 30 7 Ch in J Vet Sc i Jul. 2010 Vol. 30 No . 7 * 谭业平, 梁红茹, 牛学峰, 刘晓慧, 祝艳蕾, 郭霄峰 ( , 5 10642) : 通过分子生物学技术克隆萤火虫荧 素酶( Lucif ras ) 基因完整开放阅框后将其定向亚克隆入狂犬病病毒 HEPFlury 株全长基因组中的假基因区域, 经P CR双酶切及测序证实构建了嵌有荧 素酶基因的狂犬病病毒全 长cDNA 感染性克隆之后, 利用反向遗传操作技术进行病毒拯救, 抗核蛋白单抗直接免疫荧 染色结果显示, 成 功拯救了携荧 素酶基因的重组狂犬病病毒r HEPluc 株, RTPCR 证实插入的荧 素酶基因能够稳定遗传; 荧 素酶活性分析显示, 荧 素酶基因成功表达并具有良好生物学功能; 这为应用活体成像技术研究荧 素酶标记狂 犬病病毒在小鼠体内侵染移行规律奠定了基础 : 狂犬病病毒; 反向遗传技术; 荧 素酶 : S852. 655; R535 : A : 2010) Development of recombinant rabies virus expressing luciferase using reverse genetics * TAN Y ping, LIANG Hongru, NIU Xu f ng , LIU Xiaohui, ZHU Yanl i, GU O Xiaof ng ( Col l eg e of Vet er inar y M ed ici ne, S out h Chi na A g r ic ul tur al Univ e rs i ty , Guangz hou 510642, Ch ina) Abstract: An op n r ading fram of fir fly lucif ras was clon d and subclon d into th ps udog n r gion of th rabi s H EPFlury st rain. And th n, r combinant rabi s virus carry ing lucif ras g n ( rH EPluc) w as construct d by r v rs g n tics sy st m. Dir ct immunofluor sc nc analysis r v al d that th r combinant rabi s virus rH EPluc w as succ dly r scu d. R v rs transcription polym ras chain r action ( RTP CR) analy sis show d lucif ras g n can inh rit stably . Lucif ras assay r sult illustrat d th rH EPluc virus can xpr ss lucif ras which r tains its biologic function. This w ork lay s a good foundation for r s arch on th inf ction and m tastasis of rabi s virus in mous by v iv o imaging t chnology. Key words: rabi s virus; r v rs g n t ics sy st m; lucif ras * Cor r esp ond ing a uthor , Ema il xf g uo@scau . ed u. c n , [ 56] , , RNA