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| Construction of HEK-293T Cell Line with Stable Expression of Histone H2B and Evaluation of Proliferation Efficiency of Herpes simplex virus |
| WEI Da-Li1,2, QU Min2, KE Qun-Hua1,2, LIU Yi-Jia2, ZHANG Li-Na1,2, SUN Yue-Feng1,2, YIN Xiang-Ping2, JI Peng1,* |
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2 Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences/College of Veterinary Medicine, Lanzhou University/State Key Laboratory for Animal Disease Controland Prevention/College of Veterinary Medicine, Lanzhou 730046, China |
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Abstract Histone H2B is an important part of the nucleosome, which affects gene transcription and biological processes, and is expected to become a molecular target for disease treatment. In this study, lentiviral transfection was applied to construct an HEK-293T cell line with stable expression of histone H2B. The recombinant plasmid pLVX-mCherry-puro-H2B was constructed using the H2B gene as the target and then co-transfected with auxiliary plasmids pMD2.G and psPAX2 into HEK-293T cells for lentiviral packaging. The Lentivirus was transduced to HEK-293T cells, and a stable histone H2B-expressing cell line was obtained through purinomycin pressure screening, which was named HEK-293T-H2B cell line. The replication capacity of Herpes simplex virus (HSV) in the HEK-293T-H2B cell line was determined by qPCR and Western blot assays. The results showed that the replication of HSV was significantly weakened in the HEK-293T-H2B cell line. qPCR was used to detect the expression of downstream interferon genes in HEK-293T-H2B and wild-type HEK-293T cells infected with HSV. The results showed that the HEK-293T-H2B cell line promoted the expression of genes induced by HSV downstream typeⅠinterferon. In conclusion, the HEK-293T-H2B cell line constructed in this study can significantly prompte the typeⅠinterferon signaling pathway and inhibit the proliferation of HSV, which provides a basis for further research on the antiviral function of histone H2B.
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Received: 23 January 2025
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Corresponding Authors:
*jip@gsau.edu.cn
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