Characteristics of transcripts and changes of UL111A gene during the productive phase of infection in HCMV
Liu Xin1,2, Xie Wangkai1,2, Sun Xiangwei1,2, Chen Zhiyuan2, Shen Xian1, Xue Xiangyang2
1.Department of General Surgery, the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou 325027, China; 2.Department of Microbiology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China
Liu Xin,Xie Wangkai,Sun Xiangwei, et al. Characteristics of transcripts and changes of UL111A gene during the productive phase of infection in HCMV[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2020, 50(2): 87-91.
Abstract：Objective: To investigate the characteristics of transcripts and changes of human cytomegalovirus (HCMV) UL111A gene at different time points during the productive phase of infection. Methods: HCMV AD169 strain was used to infect human foreskin fibroblast (HFF) at various times of 12 h, 24 h, 48 h, 3 d, 5 d, 7 d, 9 d. Then RNA was extracted, reverse transcribed into cDNA, and the characteristics of UL111A transcript were detected by PCR. The amplified products were verified by cloning and sequencing to characterize the changes of UL111A transcripts at different time points. Then eukaryotic expression vector pcDNA3.1 was used to construct the three transcripts coding region sequence vectors of UL111A which were pcDNA3.1-cmvIL-10, pcDNA3.1-
LAcmvIL-10 and pcDNA3.1-unspliced; the efficient expression of the constructed coding region vectors in 293T cells was confirmed by Western blot. Results: CmvIL-10, LAcmvIL-10 and unspliced transcripts could be detected in HFF cells which infected with HCMV AD169 strain. The unspliced transcript accounted for the most, and the cmvIL-10 transcript accounted for the least. All three transcripts could be detected at 24 h and 9 d after infection, while cmvIL-10 transcript was not detected at other time points. Furthermore, the coding region sequences of the three transcripts could all be efficiently expressed in 293T cells. Conclusion: HCMV UL111A gene has three transcripts at different time points during the proliferative infection.
