CAJ | 학술논문

目的测定中国宿主动物中Amur病毒M片段全基因序列,以了解其分子特征.方法设计特异的PCR引物,用RT-PCR法分段扩增JilinAP06株M片段全长,PCR产物经克隆后进行序列测定,然后进行系统发育分析和重组分析.结果 JilinAP06株M片段全基因序列共3615个核苷酸,A+T含量为59.3%,G+C含量为40.7%,包含1个单一的开放读码框架,其最大读码框架从41～3448,由3408个核苷酸组成,共编码1135个氨基酸.序列同源分析表明,JilinAP06株与中国的人源Amur HV株同源性为96.0%～97.8%,与韩国大林姬鼠来源的Soochong HV同源性为85.6%～86.7%,与HTNV原型株76-118同源性仅为79.5%,而与其他各型病毒间的同源性均低于79.0%.氨基酸同源分析,其氨基酸序列与中国的人源株Amur HV同源性为98.1%～98.4%,与韩国大林姬鼠来源的Soochong HV同源性为96.4%～97.0%,与HTNV原型株76-118同源性为91.7%,而与其他各型病毒间的同源性均低于92.0%.系统发育分析结果显示,JilinAP06与Amur病毒构成一个相对独立的分支.结论从中国宿主动物中获得了AmurHV的M片段全基因序列.
목적 측정중국숙주동물중Amur병독M편단전기인서렬,이료해기분자특정.방법 설계특이적PCR인물,용RT-PCR법분단확증JilinAP06주M편단전장,PCR산물경극륭후진행서렬측정,연후진행계통발육분석화중조분석.결과 JilinAP06주M편단전기인서렬공3615개핵감산,A+T함량위59.3%,G+C함량위40.7%,포함1개단일적개방독마광가,기최대독마광가종41～3448,유3408개핵감산조성,공편마1135개안기산.서렬동원분석표명,JilinAP06주여중국적인원Amur HV주동원성위96.0%～97.8%,여한국대림희서래원적Soochong HV동원성위85.6%～86.7%,여HTNV원형주76-118동원성부위79.5%,이여기타각형병독간적동원성균저우79.0%.안기산동원분석,기안기산서렬여중국적인원주Amur HV동원성위98.1%～98.4%,여한국대림희서래원적Soochong HV동원성위96.4%～97.0%,여HTNV원형주76-118동원성위91.7%,이여기타각형병독간적동원성균저우92.0%.계통발육분석결과현시,JilinAP06여Amur병독구성일개상대독립적분지.결론 종중국숙주동물중획득료AmurHV적M편단전기인서렬.
Objective To study the complete sequence of M segment of Amur virus in rodents and to explore their molecular characteristics. Methods Complete M segment of Amur virus in rodent from China was amplified by RT-PCR. The purified PCR product was cloned into pGEM-T Easy vector and then sequenced. Phylogenetic analysis on multiple nucleotide sequences was performed with the Tree PUZZLE and DNAStar software. Results The full-length of its M gene comprised of 3615 nucleotides with one open reading frame (ORF) including 3408 nucleotides and encoding a protein which comprised 1135 amino acids. The ORF was located at bases 41 to 3448. The phylogenetic analysis of JilinAP06 with other hantaviruses revealed that the complete sequence of M segment of JilinAP06 strain was closely related to those Amur viruses such as B78 strain, Liu strain and H5 strain were all from the patients. The complete sequence of M segment of JilinAP06 had only 79.5% identities with the nucleotide sequence of HTNV strain 76-118. Conclusion The complete sequence on M segment of Amur virus in rodent was first time identified in this country.