Structure Analysis And Expressions Of Specific Genes In HCMV Clinical Isolates

Objective: This study was aimed to characterize the 13 genes (UL136-UL148)that have been found in the genome of the clinical isolates of HCMV. The polymorphisms and the expression of these genes were also studied. Since these 13 genes are missing in the nondisease-causing laboratory strain (AD169), this study is going to help us to understand the pathogenesis of HCMV and to predict the outcomes of different HCMV infections.Methods: The clinic isolates of HCMV were obtained from the urine sample collected from those infants with intra-uterus HCMV infection. The virus genome DNA and total RNA were extracted; According to the genome sequenceof Toledo, primers for of UL133-UL148 genes were designed and were used to amplify the complete open reading frame of these 13 genes in our 4 different clinic isolates; The resulted open reading frames of the 13 genes were cloned into pMD18-T and their sequences were confirmed by sequencingsubsequently; Bioinformatics methods were used subsequently to analyze thepolymorphisms of these genes in different stains ; The mRNA transcripts ofthese genes were detected by RT-PCR.Results: Four HCMV low passage clinical isolates were successfully isolated.As shown by PCR, all of these four strains contained UL136-UL148 ORF regions. Thirty two complete open reading frame were amplified in total andtheir sequences were submitted to GenBank(accession No.: DQ180354～DQ180391). Although these 13 genes were highly conserved in the strains analyzed, polymorphisms were found by doing sequence alignment.. The...