摘要
目的 UL141基因的结构特征及多态性对发现人巨细胞病毒(HCMV)感染致病性机制具有指导意义。文中旨在研究广州地区HCMV临床低传代株UL141基因的序列特征。方法选取2016年3月至2017年3月在广州医科大学附属广东省妇儿医院儿科就诊的10例HCMV患儿,收集患儿尿液标本,从中分离低传代临床病毒株,应用多重PCR方法鉴定,对其进行HCMV UL141基因全序列扩增、克隆、鉴定、测序,并在Gen Bank搜索HCMV UL141同源序列,行序列分析。结果UL141基因存在UL141a及UL141b 2处开放阅读框架(ORF),分别由315及1017个核苷酸构成,Gen Bank登记号分别为DQl80372和DQl80371。UL141基因全长1277 bp,编码蛋白由338个氨基酸残基组成,UL141a及UL141b开放阅读框架ORF分别为315、1017 bp,其DNA序列比较保守,存在46处位点变异,变异皆为碱基替换,无插入或缺失。HCMV UL141编码蛋白翻译后修饰位点在所有分离株中均比较保守,所有分离株UL141蛋白的等电点均在8.36~8.68之间。结论广州地区HCMV临床低传代分离株UL141基因及其编码的氨基酸序列比较保守,但仍存在一定的多态性,为进一步揭示ULl41蛋白的功能及HCMV感染的致病机制奠定了基础。
Objective The sequence characteristics and polymorphism of the UL141 gene may help find the pathogenesis of human cytomegalovirus (HCMV) infection.This study was to charac-terize the sequences of HCMV UL141 in low-passage clinical isolates in Guangzhou . Methods We collected urine samples from 10 in-fants with clinically confirmed HCMV infection in the Guangzhou are-a, isolated low-passage clinical virus strains and identified them by multiplex PCR.We performed amplification, cloning, identification and sequencing of the HCMV UL141 gene and searched the GenBank for its homologous sequences followed by sequence analysis . Results The HCMV UL141 gene was found to have 2 open reading frames( ORF) , UL141a and UL141b, composed of 315 and 1017 nucleotides and included in the GenBank with the sequence numbers of DQ180372 and DQ180371, respectively.The full length of the UL141 gene in the low-passage HCMV clinical strain (D3) was 1277 bp, the coding protein consisting of 338 amino acid residues , and the full lengths of the ORFs UL 141a and UL141b were 315 bp and 1017 bp, respectively, with relatively conservative DNA sequences .Mutations were identified in 46 sites with base substitution but no insertion or deletion .The modification sites in all the isolated strains were relatively conservative after translation of the HCMV UL 141 coding protein.The isoelectric points of the UL141 protein were 8.36-8.68 for all the clinical isolates. Conclusion Polymorphism exists in the UL141 gene and its amino acid sequences of the HCMV low-passage clinical strains isolated from infants in Guangzhou , which has shed some light on the function of the ULl 41 protein and pathogenesis of HCMV infection .
出处
《医学研究生学报》
CAS
北大核心
2017年第10期1048-1052,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金(81070516)
广东省自然科学基金(9151008901000085
2015A030313726
2016A030313788)
广东省医学科研基金(A2015133)