从摩洛哥分离出的戊型肝炎病毒5'端帽状结构和3'端序列的鉴定。
Identification of 5' capped structure and 3' terminal sequence of hepatitis E virus isolated from Morocco.
作者信息
Chen Guo-Bing, Meng Ji-Hong
机构信息
Department of Microbiology and Immunology, Southeast University School of Medicine, Nanjing 210009, Jiangsu Province, China.
出版信息
World J Gastroenterol. 2004 Jul 15;10(14):2045-9. doi: 10.3748/wjg.v10.i14.2045.
AIM
To examine 5' and 3' terminal sequences of hepatitis E virus (HEV) isolated from Morocco, to confirm 5' methylated cap structure of the genome, and to investigate whether the 3' UTR can be used to distinguish HEV genotypes instead of HEV complete genome sequence.
METHODS
RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) was employed to obtain the 5' and 3' terminal sequences of HEV Morocco strain. The 3' UTR sequence of the Morocco strain was compared with that of the other 29 HEV strains using the DNAStar software.
RESULTS
The 5' PCR product was obtained only from the RLM-RACE based on the capped RNA template. The 5' UTR of the Morocco strain had 26 nucleotides, and the 3' UTR had 65 nucleotides upstream to the polyA. The 5' UTR between HEV strains had only point mutations of nucleotides. The phylogenetic tree based on the sequences of 3' UTR was not the same as that based on the complete sequences.
CONCLUSION
The genome of HEV Morocco strain was methylated cap structure. The 3' terminal sequence can not be used for distinguishing HEV genotype for all HEV strains in place of the whole HEV genome sequence.
目的
检测从摩洛哥分离出的戊型肝炎病毒（HEV）的5'和3'末端序列，确认基因组的5'甲基化帽结构，并研究3'非翻译区（UTR）是否可用于区分HEV基因型，而非使用HEV完整基因组序列。
方法
采用RNA连接酶介导的cDNA末端快速扩增（RLM-RACE）技术获取HEV摩洛哥株的5'和3'末端序列。使用DNAStar软件将摩洛哥株的3'UTR序列与其他29株HEV的序列进行比较。
结果
仅基于加帽RNA模板通过RLM-RACE获得了5'PCR产物。摩洛哥株的5'UTR有26个核苷酸，3'UTR在polyA上游有65个核苷酸。HEV株之间的5'UTR仅有核苷酸点突变。基于3'UTR序列构建的系统发育树与基于完整序列构建的不同。
结论
HEV摩洛哥株的基因组为甲基化帽结构。3'末端序列不能替代整个HEV基因组序列用于区分所有HEV株的基因型。
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