我国登革1型病毒广州株基因组全序列测定与分析  

Determination and analysis of the complete genome of dengue 1 virus GZ/80 strain

作　　者：李晓萸[1] 耿丽卿[1] 苑锡同[1] 于曼[1] 胡志君[1] 赵卫[1] 赵月峨[1] 杨佩英[1] 秦鄂德[1] 

机构地区：[1]军事医学科学院微生物学流行病学研究所,北京100071

出　　处：《军事医学科学院院刊》2001年第3期161-165,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金资助课题 ( 39770 0 36 )

摘　　要：目的 :对引起 1980年广州登革热流行的登革 1型病毒 (GZ/ 80 )株的基因组全序列进行测定 ,为进一步了解病毒基因组结构与功能的关系 ,从分子水平探讨登革病毒的传播和流行规律及致病机制提供依据。方法 :用广州登革热流行期间从患者血清中分离的GZ/ 80株病毒感染乳鼠 ,取鼠脑制备病毒RNA。对基因组编码区分段进行RT PCR扩增 ,非编码区采用RACE法进行cDNA扩增。然后将特异产物纯化后与pGEM Teasy载体连接 ,分别进行克隆测序。结果 :GZ/ 80株基因组全长 10 735nt,5′和 3′非编码区长度分别为 94nt和 4 65nt;基因组单一的读码框架长度为 10 176nt,编码 3392个氨基酸。与登革 1型病毒 160 0 7株、WestPac74株、新加坡S2 75 / 90株和FGA/ 89株核苷酸序列的同源性分别为 94 % ,93% ,95 %和 91% ;推测的氨基酸序列的同源性分别为 98.0 % ,97.9% ,97.9%和 97.1%。结论 :GZ/ 80株基因组大小及结构与已知的登革 1型病毒其他地区分离株一致。系统发生树分析显示 ,GZ/ 80株属于第Ⅱ基因型 ,与台湾 87株、88株和泰国 80株为同一基因型。Objective:To understand the relationship between genome structure and function, the complete nucleotide sequence and the deduced amino acid sequence of the genome of dengue1 virus GZ/80 strain isolated from a patient with dengue fever in Guangzhou in 1980. Methods:Total RNA was extracted from a suckling mice brain infected by the dengue~{*2~}1 virus GZ/80 strain. Specific primers were designed according to the sequence of dengue type 1 virus Singapore S275/90 strain. The gene fragments of the encoding region of virus genome were amplified by RT~{*2~}PCR. The cDNAs of both 5~{!d~} and 3~{!d~} nonencoding regions were amplified by RACE method. And then, the amplified fragments were cloned into pGEM~{*2~}T easy vector, transformed into the competent ~{!<~}WTBX~{!=~}E.coli~{!<~}WT~{!=~} DH5~{&A~} cells. Positive clones were screened by PCR and identified by restriction enzyme digestion. The nucleotide sequences were determined by PRISMTM ABI 377 automated sequencer.~Results:The complete genome of DEN~{*2~}1 GZ/80 strain is 10 735 nucleotides in length and contains a single long open reading frame of 10 176 nucleotides, encoding a polyprotein of 3392 amino acids.Comparing it with other DEN~{*2~}1 virus strains 16007, WestPac 74,Singapore S275/90 and FGA/89, the nucleotide sequence homology was 94%,93%,95% and 91%, and the deduced amino acid sequence homology was 98% and 97.9%~{**~} and ~{*)~}97.9%~ and 97.1%, respectively. Conclusions: The genomic size and structure of GZ/80 strain were found}to be similar to those of other dengue 1 virus strains. Dendrogram of genetic relationships of 20 dengue type 1 virus strains from different geographic areas revealsed that GZ/80 strain belongs to genotype^DEN^virus strains Taiwan 87,88 and Thailand 80 are the same gene type with it. The origin of GZ/80 strain may be from our native land.

关 键 词：登革热病毒 序列测定 基因组 

分 类 号：R373.33[医药卫生—病原生物学]