Le和Tr序列对新城疫ClassⅠ毒株复制及转录水平的影响  

Effects of Le and Tr sequences of Newcastle disease virus for ClassⅠ on viral replication and transcription

作　　者：于洋[1,2] 何孔旺[1] 封振[3] 仇旭升[2] 孙英杰[2] 丁铲[2] 

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心,江苏南京210014 [2]中国农业科学院上海兽医研究所国家家禽工程技术研究中心,上海200241 [3]江苏出入境检验检疫局动植食中心,江苏南京210001

出　　处：《中国兽医科学》2013年第7期678-683,共6页Chinese Veterinary Science

基　　金：公益性行业(农业)科研专项(201003012);国家高技术研究发展计划(863)项目(2011AA10A209)

摘　　要：为了研究新城疫病毒(NDV)前导序列(Leader,Le)和尾随序列(Trailer,Tr)对病毒转录及复制水平的影响,以9a5b病毒微型基因组为平台,通过对Le和Tr序列的二级结构预测,进行了一系列的缺失、突变、替换,成功构建了6株突变型微型基因组。通过共转染、实时荧光定量RT-PCR试验表明,当Le末端9～28nt替换为Tr末端9～28nt时,并不影响病毒基因的转录复制效率,当直接缺失Le的第2个干环结构或缺失Le 25位以后序列时,显著降低了病毒基因的转录复制效率;但当缺失Tr内部42nt或26位以后的全部序列时,病毒基因的转录复制发生了终止。证明Tr序列的保守性相对于Le序列而言,在NDV复制和转录过程中发挥了更为重要的作用。In order to study the effects on the level of viral transcription and replication of the Leader （Ire） and Trailer（Tr） sequences of Newcastle disease virus（NDV）, the 9aSb minigenome was used as a platform, according to the Le and Tr sequences,to predict their RNA secondary structure. Through a series of deletion, mutation, replacement, six mutant minigenomes were constructed. Co-transfection, real-timequantitative RT-PCR experiments showed that the replacement of the Le end of 9-28 nt by Tr end of 9- 28 nt did not affect the efficiency of the viral transcription and replication. Also, direct lack of Le of the second stem-loop structure or of the Le end of the 25 nt did not significantly affect the viral transcription and replication. But when the missing Tr mid-42 nt, or end of 26 nt, the viral replication and transcription terminated. The results prove that the conservation of the Tr sequence is more important than the Le sequence in NDV replication and transcription.

关 键 词：微型基因组 前导序列 尾随序列 

分 类 号：Q789[生物学—分子生物学]