中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
5期
399-404
,共6页
陈维欣%王环宇%付士红%李铭华%刘桂芳%姜红月%王力华%王海岩%王志玉%梁国栋
陳維訢%王環宇%付士紅%李銘華%劉桂芳%薑紅月%王力華%王海巖%王誌玉%樑國棟
진유흔%왕배우%부사홍%리명화%류계방%강홍월%왕력화%왕해암%왕지옥%량국동
盖塔病毒%衣壳蛋白基因%E2蛋白基因%3'非翻译区%核苷酸序列
蓋塔病毒%衣殼蛋白基因%E2蛋白基因%3'非翻譯區%覈苷痠序列
개탑병독%의각단백기인%E2단백기인%3'비번역구%핵감산서렬
Getah virus%Capsid gene%E2 protein gene%3' untranslated region%Nucleotide sequence
目的 通过分子生物学方法研究我国山东省2008年分离的盖塔病毒(DY0824)基因组分子生物学特征.方法 应用逆转录聚合酶链反应(RT-PCR)扩增出结构区基因和3' UTR片段,连接到载体中进行序列测定,然后用Clustal X1.83和MegaAlign软件对测定的核苷酸和推测的氨基酸序列进行比较分析,用Mega4软件绘制系统发生树.结果 DY0824病毒株衣壳蛋白由804个核苷酸组成,编码268个氨基酸,与国内外其他分离株的核苷酸同源性为95.4%～99.9%,氨基酸同源性为97.4%～100%.E2蛋白全长1266个核苷酸,编码422个氨基酸,与其他株的核苷酸同源性为94.8%～99.5%,氨基酸同源性为97.6%～100%.该病毒3' UTR由401个核苷酸组成,存在3个重复序列.结论 山东省新分离盖塔病毒衣壳蛋白和E蛋白基国与该病毒原型分离株相比分别存在7个和10个氨基酸差异位点,病毒3'UTR区域存在多个核苷酸差异位点.
目的 通過分子生物學方法研究我國山東省2008年分離的蓋塔病毒(DY0824)基因組分子生物學特徵.方法 應用逆轉錄聚閤酶鏈反應(RT-PCR)擴增齣結構區基因和3' UTR片段,連接到載體中進行序列測定,然後用Clustal X1.83和MegaAlign軟件對測定的覈苷痠和推測的氨基痠序列進行比較分析,用Mega4軟件繪製繫統髮生樹.結果 DY0824病毒株衣殼蛋白由804箇覈苷痠組成,編碼268箇氨基痠,與國內外其他分離株的覈苷痠同源性為95.4%～99.9%,氨基痠同源性為97.4%～100%.E2蛋白全長1266箇覈苷痠,編碼422箇氨基痠,與其他株的覈苷痠同源性為94.8%～99.5%,氨基痠同源性為97.6%～100%.該病毒3' UTR由401箇覈苷痠組成,存在3箇重複序列.結論 山東省新分離蓋塔病毒衣殼蛋白和E蛋白基國與該病毒原型分離株相比分彆存在7箇和10箇氨基痠差異位點,病毒3'UTR區域存在多箇覈苷痠差異位點.
목적 통과분자생물학방법연구아국산동성2008년분리적개탑병독(DY0824)기인조분자생물학특정.방법 응용역전록취합매련반응(RT-PCR)확증출결구구기인화3' UTR편단,련접도재체중진행서렬측정,연후용Clustal X1.83화MegaAlign연건대측정적핵감산화추측적안기산서렬진행비교분석,용Mega4연건회제계통발생수.결과 DY0824병독주의각단백유804개핵감산조성,편마268개안기산,여국내외기타분리주적핵감산동원성위95.4%～99.9%,안기산동원성위97.4%～100%.E2단백전장1266개핵감산,편마422개안기산,여기타주적핵감산동원성위94.8%～99.5%,안기산동원성위97.6%～100%.해병독3' UTR유401개핵감산조성,존재3개중복서렬.결론 산동성신분리개탑병독의각단백화E단백기국여해병독원형분리주상비분별존재7개화10개안기산차이위점,병독3'UTR구역존재다개핵감산차이위점.
Objective To study the genome molecular characteristics of Getah virus(DY0824)which isolated in Shandong province,2008 by molecular biology methods.Methods Reverse transcriptasepolymerase chain reaction(RT-PCR)was used to amplify the structural gene and 3'UTR fragments then the RT-PCR products were inserted into PGEM-T easy to be sequenced.Computer software was used to analyze the nucleotide and deduced amino acid sequence,and draw phylogenetic trees,including Clustal X1.83 and MegaAlign and Mega4.Results The capsid protein of DY0824 consists of 804 nucleotides,encoding 268 amino acids and the full-length of E2 protein is 1266 nucleotides,encoding 422 amino acids.The nucleotide homology of the capsid protein and the E2 protein with other strains were 95.4%-99.9%and 94.8%-99.5%,and the amino acid were 97.4%-100%and 97.6%-100%.The 3'UTR of the virus include 401 nucleotides and there are three repeat sequence elements.Conclusion Compared with the prototype virus,the Getah virus isolated in Shandong province had 7 amino acid differences in capsid protein genes and 10 amino acid differences in E protein genes.The 3'UTR region had multi-nucleotide changes.
