马传染性贫血病毒驴白细胞弱毒疫苗株及其亲本驴强毒株长末端重复序列(LTR)的启动子活性比较  

Comparison of LTR-based Promoting Activity of Donkey-adapted Equine Infectious Anemia Virus with Its Parental Virus

作　　者：周家喜[1] 孔宪刚[1] 张宝山[1] 刘胜旺[1] 刘丽玲[1] 孙成群[1] 刘相东[1] 刘永刚[1] 杨威[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《病毒学报》2002年第4期342-347,共6页Chinese Journal of Virology

基　　金：国家自然科学基金项目(39770561)

摘　　要：将马传染性贫血病毒驴强毒(donkey adaptedequineinfectiousanemiavirus,D AEIAV)、马传染性贫血病毒驴白细胞弱毒(donkeyleukocyteattenuatedEIAV,DLAEIAV)及EIAVWyoming株的全长长末端重复序列(LTR)分别克隆到报告基因载体pCAT Basicvector中,获得重组质粒p D A LTR CAT、p DLA LTR CAT及p Wyo LTR CAT。将这3个重组质粒分别体外转染EIAV阴性健康驴的白细胞,比较这三者LTR启动报告基因CAT表达的基础活性和在tat pcDNA3共转染条件下激活活性的差异。结果表明:在驴白细胞中,DLAEIAVLTR启动CAT表达的活性略高于D AEIAVLTR;而与EIAVWyoming株LTR比较,DLAEIAV与D AEIAV二者LTR启动CAT表达的活性都较低。在有共转染重组表达质粒tat pcDNA3条件下,D AEIAV、DLAEIAV及EIAVWyoming株LTR起始CAT表达的活性都得到提高,分别提高了4 8倍、6 0倍和3 2倍。上述结果提示,LTR可能是体现DLAEIAV驴白细胞适应性的因素,不一定是影响其毒力减弱的根本原因。The long terminal repeats(LTRs)of donkey-adapted EIAV(D-A EIAV),donkey leukocyte attenuated EIAV(DLA EIAV)and EIAV strain Wyoming were separately cloned into pCAT-Basic vector,and the generated recombinant plasmids were designated as p-D-A-LTR-CAT,p-DLA-LTR-CAT and p-Wyo-LTR-CATThe promoting activities of three LTRs were comparatively studied in donkey leukocyte by driving chloroamphenicol acetyltransferase(CAT)expressionThe results indicated that the promoting activity of LTR from DLA EIAV was slightly higher than that of D-A EIAV,and lower than that of strain WyomingThe CAT activity was greatly increased by 48 folds driving by LTR of D-A EIAV,60 folds with DLA EIAV and 32 folds with strain Wyoming when the cells were cotransfected with recombinant plasmid containing tat of DLA EIAVThe above results suggested that LTR might play a role during DLA EIAV adaption in donkey leukocyte,but not a major determinant of its virulence

关 键 词：马传染性贫血病毒 驴白细胞弱毒疫苗株 亲本驴强毒株 长末端重复序列 启动子活性比较 

分 类 号：S852.659.3[农业科学—基础兽医学] S[农业科学—兽医学]