Book chapter
Analysis of nonessential gene function in recombinant MHV-JHM: Gene 4 knockout recombinant virus
The Nidoviruses, Vol.494, pp.83-89
Advances in experimental medicine and biology, 494, Springer
2001
DOI: 10.1007/978-1-4615-1325-4_13
PMID: 11774550
Abstract
The large size of the Coronavirus genome has made reverse genetics difficult. Targeted recombination, a technique developed by Masters and colleagues, has facilitated the introduction of mutations into the Coronavirus genome (Kuo et al., 2000). Previous work by Skinner and Siddell showed that MHV-JHM ORF 4 encodes a 15 kDa protein composed of 139 amino acids. This protein is relatively rich in threonines and includes a hydrophobic region. The N-terminus contains a potential membrane-anchoring region and the C-terminus has a possible RNA binding region (Skinner and Siddell, 1985). MHV-S, a natural variant, does not encode a functional ORF 4 suggesting that the ORF 4 product was not necessary for growth in tissue culture cells or animals. Additionally this strain contained a deletion within ORF 5a (Yokomori and Lai, 1991). Lack of mRNA 4 synthesis most likely resulted from a point mutation in the intergenic sequence (UCUAAAC to UUUAAAC). In this study, targeted recombination was used to genetically disrupt ORF 4. This recombinant virus was then analyzed in a murine model of encephalitis.
Details
- Title: Subtitle
- Analysis of nonessential gene function in recombinant MHV-JHM: Gene 4 knockout recombinant virus
- Creators
- Evelena Ontiveros - Interdisciplinary Program in Immunology, University of Iowa, Iowa City, Iowa 52242, USALili KuoPaul MastersStanley Perlman
- Resource Type
- Book chapter
- Publication Details
- The Nidoviruses, Vol.494, pp.83-89
- Series
- Advances in experimental medicine and biology; 494
- DOI
- 10.1007/978-1-4615-1325-4_13
- PMID
- 11774550
- NLM abbreviation
- Adv Exp Med Biol
- ISSN
- 0065-2598
- Publisher
- Springer; Boston, MA
- Grant note
- F31 AI10348 / NIAID NIH HHS NS 36592 / NINDS NIH HHS
- Language
- English
- Date published
- 2001
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Infectious Disease (Pediatrics)
- Record Identifier
- 9983777478102771
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