CAJ | 학술논문

目的研究广州地区人巨细胞病毒(HCMV)临床低传代分离株UL143基因的多态性.方法对3株经多重PCR鉴定的HCMV临床低传代分离株进行HCMV UL143基因全序列扩增,扩增产物克隆到pMD18-T载体上测序,并将其序列与GenBank中公布的其它临床分离株UL143基因一起进行分析.结果 D3株UL143基因因碱基缺失形成多处终止密码无法产生有功能的蛋白:Toledo株UL143基因开放读码框由279个核苷酸组成,编码蛋白由92个氨基酸残基组成;其它临床分离株UL143基因开放读码框均由252个核苷酸组成,DNA序列比较保守,变异均为碱基替换,编码蛋白由83个氨基酸残基组成,氨基酸序列也很保守,不同临床分离株氨基酸变异率为1.2%～2.4%;HCMV UL143蛋白翻译后修饰位点在除Toledo株之外的所有分离株中均高度保守,没有缺失或新增;不同临床分离株UL143蛋白二级结构有所不同;除Toledo株外,其余分离株UL143蛋白的等电点均为8.75.结论临床低传代分离株HCMV UL143基因DNA及其编码产物的氨基酸序列极为保守,但仍存在一定多态性.
목적 연구엄주지구인거세포병독(HCMV)림상저전대분리주UL143기인적다태성.방법 대3주경다중PCR감정적HCMV림상저전대분리주진행HCMV UL143기인전서렬확증,확증산물극륭도pMD18-T재체상측서,병장기서렬여GenBank중공포적기타림상분리주UL143기인일기진행분석.결과 D3주UL143기인인감기결실형성다처종지밀마무법산생유공능적단백:Toledo주UL143기인개방독마광유279개핵감산조성,편마단백유92개안기산잔기조성;기타림상분리주UL143기인개방독마광균유252개핵감산조성,DNA서렬비교보수,변이균위감기체환,편마단백유83개안기산잔기조성,안기산서렬야흔보수,불동림상분리주안기산변이솔위1.2%～2.4%;HCMV UL143단백번역후수식위점재제Toledo주지외적소유분리주중균고도보수,몰유결실혹신증;불동림상분리주UL143단백이급결구유소불동;제Toledo주외,기여분리주UL143단백적등전점균위8.75.결론 림상저전대분리주HCMV UL143기인DNA급기편마산물적안기산서렬겁위보수,단잉존재일정다태성.
Objective To investigate the polymorphism of human cytomegalovius UL143 gene of low passage clinical isolates in Guangzhou,China.Method PCR was performed to amplify the entire HCMV ULl43 gene region of 3 clinical isolates,which had been proven by multiplex PCR.The amplification products were cloned into pMD18-T-Vector and subjected to sequencing.The result of DNA sequences were analyzed together with the one of published homologous sequences in GenBank from 14 clinical isolates.Result There were several stop codons in UL143 gene due to a base deletion in open reading frame (ORF) of D3 isolate,which could lead to produce non-functional protein.UL143 ORF of Toledo isolate consisted of 279 nueleotides,encoding a protein with 92 amino acids.UL143 ORFs of other isolates consisted of 252 nueleotides,encoding a protein with 83 amino acids.The DNA sequences were quite conserved and all the variations were base substitution.The amino acid sequences of different isolates were highly conserved.with variation of 1.2%-2.4%.There were no additional or deleted sites of post translational modification of UL143 protein in all clinical isolates except Toledo isolate.There were some differences in the secondary structure among different isolates.The isoelectric point of UL143 protein of all clinical isolates except Toledo isolate was 8.75.Conclusion All DNA and deduced amino acid sequences of UL143 gene shared great similarity among HCMV clinical strains regardless of their polymorphism.