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目的研究广州地区人巨细胞病毒(HCMV)临床低传代分离株UL140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UL140基因进行全序列扩增、克隆、鉴定、测序及呈报Genbank,应用生物信息学方法分析UL140基因翻译后修饰位点、二级结构及等电点。结果成功分离3株HCMV临床病毒株(D2、D3、D52),经鉴定后被Gen Bank收录。序列号分别为:DQ180373、DQ180385和DQ180357。分析显示HCMV临床低传代D2、D3和D52病毒株的UL140基因全长576 bp,其DNA序列比较保守,编码蛋白由191个氨基酸残基组成,HCMV UL140蛋白翻译后修饰位点在所有分离株中均比较保守;所有株外,其余分离株UL140蛋白的等电点均在4.88-5.12之间。结论广州地区人巨细胞病毒UL140基因及其编码的氨基酸序列相对保守,但仍存在一定的多态性,与Toledo相比,本地区分离得到病毒株具有其明显差异,这一差异可能对HCMV临床感染与致病机制研究具有重要影响。
Objective To study the polymorphism of UL140 gene in human clinical isolates of human cytomegalovirus (HCMV) from Guangzhou. Methods Three strains of UL140 gene of HCMV clinical isolates identified by multiplex PCR were amplified, cloned, identified, sequenced and reported to Genbank. The bioinformatics methods were used to analyze the post-translational modification sites and secondary structure of UL140 gene Isoelectric point. Results Three HCMV clinical isolates (D2, D3, D52) were successfully isolated and identified by Gen Bank. Serial numbers are: DQ180373, DQ180385 and DQ180357. The results showed that UL140 gene of HCMV clinical low-passaged D2, D3 and D52 strains was 576 bp in length with a conserved DNA sequence. The encoded protein consisted of 191 amino acid residues. The HCMV UL140 protein posttranslational modification site was found in all isolates All of the isolates showed isoelectric point of 4.88-5.12. Conclusion Guangzhou human cytomegalovirus UL140 gene and its encoded amino acid sequence is relatively conservative, but there are still some polymorphisms, compared with Toledo, the strains isolated in the region has its significant difference, the difference may be HCMV clinical Infection and pathogenesis research has a significant impact.