Construction and Identification of Recombinant HeLa Cells with Human Cytomegalovirus Tegument Protein UL23 Expression

Abstract

Aim: Human cytomegalovirus (HCMV), a member of the human herpesvirus family, is a common opportunistic virus causing severe ailments and deaths in people with immature or compromised immune systems. The function of UL23, a tegument protein encoded by HCMV, is poorly understood yet. We established a HeLa cell line with stable and effective expression of HCMV pUL23 to study the situation of viral proteins in host cells in vitro.

Material and method: To generate recombinant lentiviral particles, DNAs of constructs pCDH-UL23-FLAG, psPAX2,and pMD2.G were co-transfected into Lenti-X 293T cells. The stably pUL23-expressing HeLa cells resistant to puromycin were obtained after recombinant lentivirus infection.The UL23 gene was successfully amplified through RT PCR in DNA extraction from the cell line and the expression of pUL23 was identified through Western blotting.

Results and discussion: The results verified that a HeLa cell line stably expressing pUL23 was established with a lentivirus expression system.

Conclusion: The HeLa cell line with stable and effective expression of HCMV pUL23 paves the way for future study on biological functions and mechanism of protein UL23.