K亚群禽白血病病毒5′LTR序列及启动活性分析  

The Research of 5′LTR Promoter Activity of Avian Leukosis Virus Subgroup K

作　　者：赵子君 饶明章 陈建 张杰 袁丽霞 廖明 曹伟胜 ZHAO Zi-jun;RAO Ming-zhang;CHEN Jian;ZHANG Jie;YUAN Li-xia;LIAO Ming;CAO Wei-sheng(Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, Key Labora tory of Zoonoses Control and Prevention of Guangdong , College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China)

机构地区：[1]华南农业大学兽医学院农业部兽用疫苗创制重点实验室广东省动物源性人兽共患病预防与控制重点实验室,广州510642

出　　处：《畜牧兽医学报》2018年第4期754-760,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31672552);广东省自然科学基金(2016A030313403);广东省家禽产业技术体系疾病控制岗位专家(2016LM1114);国家肉鸡产业技术体系(CARS-41-G16)

摘　　要：通过对ALV-K 5′LTR序列及其启动活性比较分析,以探究5′LTR对ALV-K体外复制能力的影响。选取广东某黄羽祖代种鸡场健康鸡群中分离到的ALV-K(GDFX0601)毒株作为研究对象,将其5′LTR核苷酸序列与国内外不同亚群ALV分离株比较分析,并将该ALV-K接种DF-1细胞,采用ELISA对细胞培养上清进行ALV p27抗原检测以分析其在DF-1细胞上的复制能力,还分别将ALV-K(GDFX0601)、ALV-J(CHN06)、ALV-E(ev-1)和ALV-K(JS11C1)5′LTR克隆进pGL3-Basic载体,分别转染DF-1、CEF和293T细胞后测定荧光素酶活性来评估不同毒株5′LTR的启动活性。结果显示,ALV-K(GDFX0601)与其他内源性ALV LTR相似性为98.5%,而与其他外源性ALV的相似性仅有70%左右,且其LTR U3区转录调节元件与外源性病毒差异较大,与内源性病毒相似。ALV-K(GDFX0601)在ALV易感细胞DF-1上的复制速度与感染能力均弱于外源性病毒ALV-A、ALV-B和ALV-J。5′LTR的启动活性试验发现ALV-K(GDFX0601)毒株LTR的启动活性比ALV-J和LTR为外源性的ALV-K(JS11CI)毒株的启动活性弱(P<0.05)。ALV-K比其他亚群ALV复制能力低可能与5′LTR启动活性有关。In this study,we investigated the proliferation of avian leukosis virus subgroup K（ALV-K）and the activity of LTR.An ALV-K strain was used to infect DF-1 cells and the LTR was cloned into pGL3-Basic vector.The replication ability of the virus on ALV susceptible cell DF-1 was evaluated by ALV p27 antigen ELISA,and the promoter activities of the LTR of ALV-K,ALV-E and ALV-J strains were measured using a Dual-Glo Luciferase Assay System.The results showed that promoter activity of ALV-K LTR was significantly lower than those of other exogenous ALV in DF-1,CEF and 293 Tcells.The relatively poorer replication ability of ALV-K may be related to its weak promoter activity of the LTR.

关 键 词：ALV-K 致病性 LTR 启动活性 

分 类 号：S852.659.3[农业科学—基础兽医学]