马胞嘧啶脱氨基酶A3Z3的多态性及抗马传染性贫血病毒作用  

The polymorphism of equine cytosine deaminase A3Z3 and its anti-equine infectious anemia virus activity

作　　者：张泽力[1,2] 马建 陈方锐 尹鑫[2,3] 曹苏亚[1,2] 艾有为[1,2] 王玉龙[1] 王晓钧[2] 

机构地区：[1]东北林业大学野生动物资源学院,黑龙江哈尔滨150040 [2]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/大动物研究室,黑龙江哈尔滨150001 [3]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2014年第6期427-430,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：中央高校基本科研业务费用专项资金资助(DL13CA04);国家自然科学基金(31072113);兽医生物技术国家重点实验室自主课题(SKLVBP201304);中央级公益性基本科研业务费预算增量项目(2012ZL080);黑龙江自然科学基金(C201116)

摘　　要：马胞嘧啶脱氨基酶家族成员之一的A3Z3分子(eqA3Z3),可通过胞嘧啶脱氨基酶活性限制马传染性贫血病毒(EIAV)的复制。为研究eqA3Z3分子多态性与其抗EIAV活性之间的关系,本实验采用RT-PCR技术从10匹马体内扩增A3Z3基因,对获得的不同马体内A3Z3分子的序列进行比对分析,发现了两种不同于已报道eqA3Z3分子的突变体。将其克隆于真核表达载体pcDNA-HA中,并分别与EIAV感染性质粒共转染293T细胞,通过GST Pull-down方法证明了3种分子均能够与EIAV Gag蛋白相互作用。此外,western blot结果显示3种分子均能够被包装进病毒粒子。同时荧光素酶活性结果表明3种分子具有相同的限制EIAV复制功能。以上研究结果有助于进一步深入了解和评价马属动物Apobec3分子抗逆转录病毒的作用。Equine A3Z3, one of the members of cytosine deaminase family, has the ability of inhibiting the replication of equine infectious anemia virus （EIAV） by cytosine deaminase activity. To investigate the relationship between eqA3Z3 polymorphisms and their antiviral activity, eqA3Z3 genes were amplified from ten horses by RT-PCR. Then the sequences of eqA3Z3 were analysed, and two new mutants of eqA3Z3 genes were identified. These three genes were inserted into the eukaryotic expression vector pcDNA-HA. The interaction between eqA3Z3 and EIAV Gag was detected by GST Pull-down which was followed by co-transfecting the recombinant eqA3Z3 plasmids and EIAV infectious plasmid into 293T cells, the result revealed that three types of proteins were able to interact with EIAV Gag. Additionally, the result of western blot showed that three eqA3Z3 were also able to be packaged into virions. Furthermore, the anti-EIAV result revealed that three eqA3Z3 harbored the similar anti-EIAV activities. This experiment contributed to further study of antiretroviral activities of equine Apobec3.

关 键 词：马A3Z3 多态性 马传染性贫血病毒 抗病毒作用 

分 类 号：S852.65[农业科学—基础兽医学]