中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2009年
4期
244-249
,共6页
王波%李月琴%胡兢晶%苏海浩%丁俊彩%田传军%张纯青%周天鸿
王波%李月琴%鬍兢晶%囌海浩%丁俊綵%田傳軍%張純青%週天鴻
왕파%리월금%호긍정%소해호%정준채%전전군%장순청%주천홍
巨细胞病毒%多态性%单核苷酸%基因%UL136%临床病毒株%生物信息学
巨細胞病毒%多態性%單覈苷痠%基因%UL136%臨床病毒株%生物信息學
거세포병독%다태성%단핵감산%기인%UL136%림상병독주%생물신식학
Cytomagalovirus%Polymorphism%single nucleotide%Genes%UL136%Clinical isolates%Bioinformatics
目的 研究广州地区新生儿感染人巨细胞病毒(HCMV)临床低传代株UL136基因的序列特征与基因多态性.方法 从10例广州感染新生儿体内分离获得2株(D2、D3)临床HCMV分离株,经多重PCR鉴定后进行UL136基因全序列扩增.PCR产物纯化后进行基因克隆,构建HCMV UL136-pMD18-T重组质粒.经基因测序及应用生物信息学分析方法 ,分析其核酸序列稳定性、编码蛋白质的二级结构与特征.结果 成功分离2株HCMV临床分离株,测序结果 显示,D2、D3及与GenBank中公布的11株临床分离株(4J、51C、39J、33J、63J、22M、10J、32C、29C、27C、Toledo)中,UL136序列高度保守.同源性分析显示在UL136全基因序列1019个核苷酸中,存在30个位点变异,所有的变异均为碱基替换,无插入及缺失突变.编码蛋白的氨基酸序列也高度保守,240个氨基酸残基中,不同临床分离株氨基酸变异率为1.6%～3.7%.不同分离株的UL136蛋白中参与形成二级结构的氨基酸数目及等电点不同.进化树分析结果 显示D2和D3均属于1a群.结论 广州地区临床低传代分离株HCMV UL136基因核苷酸序列及其氨基酸序列极为保守,但仍存在一定多态性.其基因的稳定性提示HCMV UL136开放阅读框(ORF)可能是一个具有重要功能的基因.其编码后修饰位点提示UL136可能与膜受体介导的细胞信号转导通路有关.
目的 研究廣州地區新生兒感染人巨細胞病毒(HCMV)臨床低傳代株UL136基因的序列特徵與基因多態性.方法 從10例廣州感染新生兒體內分離穫得2株(D2、D3)臨床HCMV分離株,經多重PCR鑒定後進行UL136基因全序列擴增.PCR產物純化後進行基因剋隆,構建HCMV UL136-pMD18-T重組質粒.經基因測序及應用生物信息學分析方法 ,分析其覈痠序列穩定性、編碼蛋白質的二級結構與特徵.結果 成功分離2株HCMV臨床分離株,測序結果 顯示,D2、D3及與GenBank中公佈的11株臨床分離株(4J、51C、39J、33J、63J、22M、10J、32C、29C、27C、Toledo)中,UL136序列高度保守.同源性分析顯示在UL136全基因序列1019箇覈苷痠中,存在30箇位點變異,所有的變異均為堿基替換,無插入及缺失突變.編碼蛋白的氨基痠序列也高度保守,240箇氨基痠殘基中,不同臨床分離株氨基痠變異率為1.6%～3.7%.不同分離株的UL136蛋白中參與形成二級結構的氨基痠數目及等電點不同.進化樹分析結果 顯示D2和D3均屬于1a群.結論 廣州地區臨床低傳代分離株HCMV UL136基因覈苷痠序列及其氨基痠序列極為保守,但仍存在一定多態性.其基因的穩定性提示HCMV UL136開放閱讀框(ORF)可能是一箇具有重要功能的基因.其編碼後脩飾位點提示UL136可能與膜受體介導的細胞信號轉導通路有關.
목적 연구엄주지구신생인감염인거세포병독(HCMV)림상저전대주UL136기인적서렬특정여기인다태성.방법 종10례엄주감염신생인체내분리획득2주(D2、D3)림상HCMV분리주,경다중PCR감정후진행UL136기인전서렬확증.PCR산물순화후진행기인극륭,구건HCMV UL136-pMD18-T중조질립.경기인측서급응용생물신식학분석방법 ,분석기핵산서렬은정성、편마단백질적이급결구여특정.결과 성공분리2주HCMV림상분리주,측서결과 현시,D2、D3급여GenBank중공포적11주림상분리주(4J、51C、39J、33J、63J、22M、10J、32C、29C、27C、Toledo)중,UL136서렬고도보수.동원성분석현시재UL136전기인서렬1019개핵감산중,존재30개위점변이,소유적변이균위감기체환,무삽입급결실돌변.편마단백적안기산서렬야고도보수,240개안기산잔기중,불동림상분리주안기산변이솔위1.6%～3.7%.불동분리주적UL136단백중삼여형성이급결구적안기산수목급등전점불동.진화수분석결과 현시D2화D3균속우1a군.결론 엄주지구림상저전대분리주HCMV UL136기인핵감산서렬급기안기산서렬겁위보수,단잉존재일정다태성.기기인적은정성제시HCMV UL136개방열독광(ORF)가능시일개구유중요공능적기인.기편마후수식위점제시UL136가능여막수체개도적세포신호전도통로유관.
Objective To investigate the nucleotide sequence characterization and genetic polymorphism of human cytomegalovirus (HCMV) UL136 gene in low-passage clinical isolates of infants in Guangzhou. Methods Two clinical HCMV strains (D2 and D3) were isolated from 10 infected infants in Guangzhou. After identification by multiplex PCR, the entire HCMV UL136 gene sequence was amplified. The pured PCR products were cloned into pMD18- T-vector to construct HCMV UL136-pMD18- T recombinant plasmid. The sequence stability, secondary structure and characterization of coding protein were analyzed by sequencing and biological methods. Results Two HCMV clinical strains were successfully isolated. After cloning and sequencing, UL136 genes of D2, D3 and 11 clinical isolates published in GenBank (4J, 51C, 39J, 33J, 63J, 22M, 10J, 32C, 29C, 27C, Toledo) were shown to be highly conservative. Homology analysis revealed mutations in 30 sites among 1019 base pairs in UL136 gene sequence, which were base substitutions without insertion and deletion. High conservative amino acid sequence was also seen in coded proteins. Among 240 amino acid residues, the aberration rates across various clinical isolates ranged from 1.6% to 3.7%. The UL136 properties among these isolates differed in terms of amino acid amounts involved in secondary structure and their isoelectric points. Cladogram showed that D2 and D3 were members of group la. Conclusion All nucleotide and deduced amino acid sequences of UL136 gene share great conservation among low passage HCMV clinical strains in Guangzhou regardless of their polymorphism. The gene stability implies that UL136 open reading frame of HCMV plays an important role in viral infection. The post-translation modified sites suggest that UL136 protein may be related to membrane-receptor mediated cellular signal transduction.
