The Expression and Anti-apoptotic Function of Herpes Simplex Virus Type 2 Latency Associated Transcript-RL1

Abstract

Abstract: It was to study the expression of herpes simplex virus type 2 latency-associated transcript(LAT)RL1 and its anti-apoptosis function induced by actinomycin D in vero cells. The recombinant plasmid pEGFP-ORF1 was constructed and transfected into Vero cells, expression was determined by green fluorescent protein and RT-PCR .The changes of Vero cells morphology induced by Actinomycin D were observed by Hochest33342 fluorescence staining, cells apoptosis rate was detected by flow cytometry . Membrane potential JC-1 fluorescence was observed, and Caspase 3 detected of apoptosis protein activity. Results showed that the green fluorescent protein has a high expressed efficiency in Vero cells, and target gene was detected by RT-PCR. Hochest33342 staining reavealed that Vero cells transfected with pEGFP-RL1 and induced apoptosis by actinomycin D had no changes in morphology. Flow cytometry assay showed that the cells apoptosis rate were no significant difference between pEGFP-RL1group and the normal group, but the cells apoptosis rate of pEGFP-RL1 was remarkable lower than the pEGFP-C2 group. Transfected with the recombinant plasmid pEGPF-RL1 cells by JC-1 staining, the proportion is much larger than red cells green cells transfected with empty vector pEGFP-C2. Caspase-3 results indicate that induction of apoptosis by transfected with empty vector pEGFP-C2 Vero cells, and the activity was significantly higher than those transfected with pEGFP-RL1 by actinomycin D-induced apoptosis of Vero cells and normal cells. HSV-2 LAT RL1 gene can effectively expressed in Vero cells and can protect Vero cells from apoptosis induced by actinomycin D.

Key words: Herpes simplex virus, LAT, RL1, Actinomycin D, Apoptosis,Vero cells

Gao Ruidi, Yang Huilan, Zhong Feifei, Lü Fangbiao. The Expression and Anti-apoptotic Function of Herpes Simplex Virus Type 2 Latency Associated Transcript-RL1[J]. Biotechnology Bulletin, 2014, 0(3): 187-192.

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