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Title: Construction of a herpes simplex virus type 1 derived vector for targeted long term gene expression in the mammalian nervous system
Author: Cai, S.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 1999
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The herpes simplex virus type 1 (HSV -1) latency-associated transcripts (LATs) consist of a series of nuclear RNAs which are transcribed antisense to part of the ICP0 gene. Authentic LAT promoter (LAP) activity is responsible for the continued transcription of LATs in latently infected neurones. In this study, an analysis of regions encompassing LAP which contain regulatory elements important for long term LAP activity has been undertaken. Insertion of a reporter gene cassette, consisting of an EMCV (encephalomycarditis virus) IRES (internal ribosomal entry site) linked lacZ gene (IRES-lacZ) 77bp downstream of the HSV-1 LAT transcription start site showed that this recombinant virus (LβC) expressed β-gal in a small number of neurones in acutely infected mouse cervical ganglia and that a decrease in the number of β-gal positive neurones was observed at late time points. However, a virus designated LβA which has an IRES-lacZ cassette inserted 1.5kb downstream of the LAT transcription start site, resulted in a greater number of neurones expressing β-gal at day 30 postinfection (p.i.) than at day 5 p.i. of mice. To define the cis-acting elements required for long term LAP activity, a 3.3 kb fragment encompassing LAP was utilised to drive expression of an IRES-lacZ cassette inserted at the non-essential Us5 locus of a parental virus lacking both copies of this promoter region. This virus (Us5HβG) expressed β-gal in latently infected neurones analysed at day 30 p.i. and the amount of β-gal expression in neurones increased during the transition between the acute day 5 p.i. and day 30 p.i. time points. Analysis of ganglia latently infected with either LβA or Us5HβG at day 56 p.i. revealed a decrease in the number of β-gal expression neurones as compared to the day 30 p.i. time point, although the intensity of β-gal signal remained unchanged.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available