我国登革4型病毒广州株基因组非编码区序列测定及分析  

DETERMINATION AND ANALYSIS OF THE SEQUENCES OF THE NONCODING REGIONS OF DENGUE TYPE 4 VIRUS GRANGZHOU STRAIN ISOLATED IN CHINA

作　　者：王鹏程[1] 秦鄂德[1] 耿丽卿[1] 于曼[1] 赵卫[1] 陈水平[1] 苑锡同[1] 

机构地区：[1]军事医学科学院微生物流行病学研究所,北京100071

出　　处：《中国人兽共患病杂志》2001年第4期6-9,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助项目 (39770 0 36 )

摘　　要：目的　测定和分析我国登革 4型病毒广州 (D4-B5 )株基因组非编码区 (NCR)序列 ,为探讨其结构特征与功能的关系提供依据。方法　从登革 4型病毒感染的乳鼠脑中提取总RNA ,采用RACE法扩增病毒基因组 5’和 3’端片段 ,分别将其克隆至pGEM -T载体 ,挑取阳性克隆进行序列测定 ;利用RNAdraw软件对非编码区二级结构进行预测。结果与结论　我国登革 4型病毒广州株基因组 5′和 3′非编码区长度分别为 10 1nt和 40 3nt,具有黄病毒共有的保守序列和二级结构。与登革4型病毒多米尼加分离株比较显示 ,5′NCR第 5 7和 36位核苷酸的改变对其二级结构有显著的影响 ;3′NCR多处核苷酸差异导致两者预测的二级结构差别较大 ,但均能形成相同的保守结构 ,它们可能对病毒基因组RNA的复制。Aim To determine and analyse the sequences of the noncoding regions(NCR)of dengue type 4 virus Guangzhou strain(D4-B5)isolated in China and provide information about the relationship between viral NCRs and their functions.Methods With total RNA extracted from the brain of suckling mouse inoculated with D4-B5,the specific fragments of viral 5' and 3' NCRs were amplified by RACE and cloned into pGEM-T vectors,respectively.Positive clones were identified and the inserted fragments were sequenced.The secondary structures of 5'and 3' NCR were predicted with RNAdraw software,respectively.Results and Conclusions The 5' and 3' NCRs of D4-B5 strain are 101nt and 403nt in length respectively and have the commonly-conserved sequences and secondary structures of Flavivirus.Comparison with isolate from Dominica (D4-814669)suggests that(i:The differences at nucleotides 57 and 36 of 5' NCR have significant influence on the conformation of the predicted secondary structure;(ii:Although with many differences,the two strains have the same conserved structures which are of great importance to the replication and package of virus genome RNA.

关 键 词：登革4型病毒 基因组非编码区 序列测定 分析 

分 类 号：R373.33[医药卫生—病原生物学]