将人类巨细胞病毒HCMVTowne株接种至人包皮成纤维细胞(HFF)中,收集病毒感染后的细胞,抽提病毒RNA,运用RT-PCR技术扩增基因的序列,对PCR产物作TA克隆,重组体进行测序和生物信息学分析。结果表明,HCMV的UIA9区域存在一个新转录本,暂命名为UIA9X。将所获得的序列与HCMV基因组比对分析表明,UIA9X序列为68783bp到78608bp处的基因片段,并发现从69194bp到71530bp处有个典型的内含子。
Incubated HCMV Towne to the well growing HFF cells; Collected the ceils while the HFF cells appearit~g eytopathic effects; Extracted the HCMV RNA from the infected ceils. Amplified the cDNA of the gene by RT-PCR. Finally, the production of RT-PCR were cloned into pMD18-T vectors and sequenced subsequently. And the sequence of ULA49X was analyzed by bioinfonnatics. Our research show that, there is a new gene in the UL4-9 region of HCMV which we called it ULA9X. The sequence of UL49X gene was from 68 783 bp to 78 608 bp, and we found that there was a intron between 69 194 bp and 71 530 bp.