Abstract
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. Key words: Cytomagalovirus; Polymorphism; single nucleotide; Genes; UL136; Clinical isolates; Bioinformatics
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