Establishment And Characterization Of A Reference Strain Of Herpes Simplex Virus Type 1

Herpes simplex virus (HSV) type 1 is a member of the herpesviridae family, the virus frequently infects human beings,causing a range of diseases,including gingival stomatitis;encephalitis;corneal conjunctivitis. HSV1 can productively infect many different types of cells and establish latent infections in neuronal ganglia in vivo.There is no vaccine against HSV infection and complete removal of the virus drug is developed. In addition,HSV is characterized by large cyclic DNA and latent infection,the HSV-vectors are especially useful for delivery to the nervous system.In this study,a strain of virus was isolated from blisters of patient use Vero,and the isolated virus is proved to be HSV-1 through Sequence analysis and Electron microscopy identification,named HSV1-ZW6 strain.For proliferative rule of HSV1-ZW6,including the infection of different cell line,one step growth curve drawing,mouse neurovirulence test etc.The results show that the neurovirulence of this isolated strain higher than the passage strain HSV1-8F.The genome of this virus are completed by high-throughput sequencing technologise and repairing with Sanger method,for gene prediction and annotation to obtain the structure graph of genome,completing phylogenetic tree based on parts of gene,then found a special 32bp repeat sequence in IRL/TRL based on analysis of sinple sequence repeats. According to the known sequence,a recombinant virus ZW6-EGFP was successfully obtained via CRISPR-Cas9 system. In addition, to studing KMB17 cells generation and changes in telomere length, a diploid cell model is established.Then,at the cellular level to detect the KMB17 cells infect with herpes simplex virus, preliminary findings HSV1 infect KMB17 can effect the telemore length, and cancer cells or immortalized cell lines is not found similar results.Through the isolation,identification and complete genome sequencing of HSV1, this study establishes a strain of HSV 1 reference strain and its report virus.We just have a preliminary exploration of HSVl-ZW6,there are many worthy of study above this virus. The study is only a preliminary exploration on the interaction between virus and cell, the reference strains of biological characteristics needs to be more studied further...