Replication characteristics of herpes simplex virus type-1 (HSV-1) recombinants in 3 types of tissue cultures

Karimi, A. and MacLean, A. (2005) Replication characteristics of herpes simplex virus type-1 (HSV-1) recombinants in 3 types of tissue cultures. Iranian Biomedical Journal, 9 (3). pp. 95-101.

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Abstract

A complication in the analysis of the role of ICP34.5 gene in the herpes simplex virus type-1 (HSV-1) lifecycle is the presence of overlapping antisense gene, open reading frame P (ORF P), which is also deleted in HSV-1 ICP34.5 negative mutants. A HSV-1 wild type strain (17+) ICP34.5/ORF P deletion mutant (1716) is totally avirulent in animal models and impaired in a number of in vitro functions: replication in 3T6 cells; and replication and shutoff of cellular protein synthesis in SK-N-SH cells. To attribute characteristics of 1716 to each of these two genes (ICP34.5 or ORF P), a number of HSV-1 recombinant viruses that express ICP34.5 and ORF P independently were constructed, purified and characterized. The parent of these recombinants is 1716 and they are (so called): 1622, expressing ICP34.5; 1624/24.5, expressing ORF P; and 1625, expressing both ICP34.5 and ORF P in separate loci. Using homologous recombination in cell culture, the recombinants were constructed and their DNA were analyzed by Southern-blotting. Expression of ICP34.5 and ORF P from the recombinants was checked by Western-blotting. Using cell culture, titration and plaque assay techniques, replication kinetics of the recombinants were compared with 17+ and 1716 in 3 cell lines, BHK, 3T6, and SK-N-SH. The results showed that (i) ICP34.5 restored the ability to replicate and prevent host shutoff similar to wild type in SK-N-SH cells; (ii) ICP34.5 restored the replication phenotype to near wild type levels in 3T6 cells; and (iii) ORF P expression had no effect on the replication of these mutants. The characteristics of 1716 is obviously due to the lack of ICP34.5 and ORF P has no role in the characteristics studied. Thus, the function of ORF P still has to be determined.

Item Type: Article
Additional Information: cited By
Uncontrolled Keywords: infected cell protein 8, animal cell; animal tissue; article; cell culture; controlled study; DNA determination; gene expression; Herpes simplex virus 1; homologous recombination; human; human cell; human tissue; mouse; nonhuman; tissue culture; titrimetry; virus characterization; virus culture; virus recombinant; virus replication, Animalia; Human herpesvirus 1
Subjects: WC Communicable Diseases > Virus diseases
QU Biochemistry
Divisions: Faculty of Medicine > Basic Sciences Academic Groups > Department of Immunology
Faculty of Medicine > Basic Sciences Academic Groups > Department of Microbiology
Depositing User: zahra bagheri .
Date Deposited: 21 Aug 2017 09:29
Last Modified: 21 Aug 2017 09:29
URI: http://eprints.skums.ac.ir/id/eprint/3456

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