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作 者:周家喜[1] 张宝山[1] 刘永刚[1] 孔宪刚[1] 刘胜旺[1] 孙成群[1] 刘相东[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,黑龙江哈尔滨150001
出 处:《中国兽医学报》2002年第2期128-130,共3页Chinese Journal of Veterinary Science
基 金:国家自然科学基金资助项目 ( 39770 5 6 1)
摘 要:以马传染性贫血病毒 (EIAV)疫苗毒感染驴巨噬细胞培养物 ,在病毒繁殖期提取细胞总 RNA,反转录后使用特异引物扩增病毒基因组拼接产物。将扩增产物克隆、测序并与基因组全序列比较 ,确定了编码 EIAV反式激活蛋白(TAT)的转录产物及阅读框架 ,发现至少有 2种拼接产物编码 TAT。比较 2种阅读框架 ,以保守序列作为模板扩增得到完整的编码 EIAV反式激活蛋白的基因。The donkey macrophage culture was infected with donkey leukocyte attenuated EIAV.At the early stage of virus replication the total RNA was extracted from infected cells.After reverse transcription,polymerase chain reaction was performed with EIAV specific primers.Then PCR products were cloned in the pUC19 plasmid vector and the inserts were sequenced.Two clones encoding TAT were obtained in this study.ORF encoding TAT and splicing sites were also determined.The whole sequence of tat gene has been obtained by further polymerase chain reaction during which more conservative sequence was used as template.
关 键 词:马传染性贫血病毒 反式激活蛋白 拼接产物 测序 TAT 基因 分子克隆 序列
分 类 号:S852.652[农业科学—基础兽医学]
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