Full-Length Cloning Of HCMV UL49Z Gene And Bioinformatical Analysis

Objective:To obtain the full length cDNA of HCMV UL49Z gene by RACE technology in HCMV Towne. Using bioinformatics methods to analyse UL49Z gene's function. Setting up foundation for further research of the structures and functions of the genes in HCMV UL49 region, and also making a basic research for the molecular mechanism of the crucial region UL49.Methods:Incubated HCMV Towne to the well growing HFF cells; Collected the cells while the HFF cells appearing cytopathic effects; Extracted the HCMV RNA from the infected cells. We confirmed the position about the 3'end and the 5'end of UL49Z by using RACE and Amplified the cDNA of HCMV UL49Z gene by PT-PCR. Finally, the production of RACE and RT-PCR were cloned into pMD18-T vectors and sequenced subsequently. and the sequence of UL49Z was analyzed by bioinformatics.Results:HFF cells were successfully infected by HCMV Towne, and the HCMV total RNA was successfully extracted. We discovered a new gene by RACE and RT-PCR, we temporarily called it UL49Z. And the product of RACE and RT-PCR was successfully sequenced. We found that the full-length sequence of UL49Z is from 68629bp to 72830bp, and there is a intron between 71181bp and 71276bp.Conclusion:There is a new gene in the HCMV UL49 region. Our work setting up foundation for the further research of the structures and functions of UL49Z.