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Cloning and fine mapping the DNA of equine herpesvirus type one defective interfering particles.
Abstract: Equine herpesvirus type one (EHV-1) defective interfering (DI) particle DNA fragments were inserted into the XbaI site of the plasmid vector pACYC184. Five DI XbaI fragments, which ranged in molecular weight from 4.5 to 6.7 MDa, were selected for detailed analysis. Each DI DNA clone was labeled with 32P-deoxynucleotides by nick translation and hybridized to genomic digests of EHV-1 standard (STD) DNA bound to nitrocellulose. All five clones were shown to hybridize to DNA sequences derived from the left terminus (0.0-0.04 map units) of the long (L) region and from the short (S) region inverted repeats (IRs, 0.79-0.86 and 0.93-1.00 map units) of the STD EHV-1 genome. Restriction enzyme mapping studies and Southern blot hybridizations employing cloned STD virus DNA fragments as probes revealed that these EHV-1 DI clones contain two major domains: (1) an L terminal region which maps to 0.01-0.04 map units and is highly conserved among all five clones, and (2) a region homologous to the IRs which appears to vary between individual clones.
Publication Date: 1986-09-01 PubMed ID: 3016989DOI: 10.1016/0042-6822(86)90022-xGoogle Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research revolves around the detailed analysis of the DNA of defective interfering particles of Equine herpesvirus type one (EHV-1). The results indicate the presence of two major domains in the DNA of these particles, with variation observed among individual clones.
Research Objectives and Methods
- The primary aim of this research was to clone and create a fine map of the DNA of equine herpesvirus type one (EHV-1) defective interfering (DI) particles.
- The researchers inserted DNA fragments from EHV-1 DI particles into a specific site (XbaI site) of the plasmid vector pACYC184. These fragments were then selected for comprehensive analysis.
- The researchers used radioactive phosphorus (32P-deoxynucleotides) to label each clone and then hybridized it to genomic digests of EHV-1 standard DNA bound to nitrocellulose. This process allowed them to identify the regions of the genome where the clones associated.
Results
- All five analyzed clones were shown to hybridize to DNA sequences found at the extreme left of the long (L) region (0.0-0.04 map units) and the inverted repeats (IRs) present in the shorter (S) region (0.79-0.86 and 0.93-1.00 map units) of the standard EHV-1 genome.
- Restriction enzyme mapping studies and Southern blot hybridizations further added to this data, helping the researchers associate cloned DNA fragments from the standard virus as probes.
- These studies revealed that the cloned EHV-1 DI particles have two major domains. The first, a highly conserved region located at the terminal of the L region (0.01-0.04 map units), and the second, a region corresponding to the IRs, where differences among individual clones were observed.
Conclusion and Implications
- By mapping the DNA of EHV-1 DI particles, the study has furthered our understanding of their structure and how they function.
- These findings may be crucial in further research related to EHV-1, particularly in efforts to modify or control this virus. A detailed knowledge of the DNA structure of EHV-1 DI particles could potentially lead to the development of improved treatment strategies for diseases caused by this virus.
Cite This Article
APA
Baumann RP, Staczek J, O'Callaghan DJ.
(1986).
Cloning and fine mapping the DNA of equine herpesvirus type one defective interfering particles.
Virology, 153(2), 188-200.
https://doi.org/10.1016/0042-6822(86)90022-x Publication
Researcher Affiliations
MeSH Terms
- Cell Transformation, Neoplastic
- Cloning, Molecular
- DNA Replication
- DNA, Viral / analysis
- Defective Viruses / genetics
- Herpesviridae / genetics
- Herpesvirus 1, Equid / genetics
- Nucleic Acid Hybridization
- Repetitive Sequences, Nucleic Acid
- Sequence Homology, Nucleic Acid
- Viral Interference
- Virus Replication
Grant Funding
- AI 21996 / NIAID NIH HHS
- AI 22001 / NIAID NIH HHS
- AI 22894 / NIAID NIH HHS
Citations
This article has been cited 12 times.- Charvat RA, Zhang Y, O'Callaghan DJ. Deletion of the UL4 gene sequence of equine herpesvirus 1 precludes the generation of defective interfering particles. Virus Genes 2012 Oct;45(2):295-303.
- Ahn BC, Kim S, Zhang Y, Charvat RA, O'Callaghan DJ. The early UL3 gene of equine herpesvirus-1 encodes a tegument protein not essential for replication or virulence in the mouse. Virology 2011 Nov 10;420(1):20-31.
- Charvat RA, Breitenbach JE, Ahn B, Zhang Y, O'Callaghan DJ. The UL4 protein of equine herpesvirus 1 is not essential for replication or pathogenesis and inhibits gene expression controlled by viral and heterologous promoters. Virology 2011 Apr 10;412(2):366-77.
- Ebner PD, Kim SK, O'Callaghan DJ. Biological and genotypic properties of defective interfering particles of equine herpesvirus 1 that mediate persistent infection. Virology 2008 Nov 10;381(1):98-105.
- Ebner PD, O'Callaghan DJ. Genetic complexity of EHV-1 defective interfering particles and identification of novel IR4/UL5 hybrid proteins produced during persistent infection. Virus Genes 2006 Jun;32(3):313-20.
- Chen M, Harty RN, Zhao Y, Holden VR, O'Callaghan DJ. Expression of an equine herpesvirus 1 ICP22/ICP27 hybrid protein encoded by defective interfering particles associated with persistent infection. J Virol 1996 Jan;70(1):313-20.
- Harty RN, Caughman GB, Holden VR, O'Callaghan DJ. Characterization of the myristylated polypeptide encoded by the UL1 gene that is conserved in the genome of defective interfering particles of equine herpesvirus 1. J Virol 1993 Jul;67(7):4122-32.
- Harty RN, Colle CF, Grundy FJ, O'Callaghan DJ. Mapping the termini and intron of the spliced immediate-early transcript of equine herpesvirus 1. J Virol 1989 Dec;63(12):5101-10.
- Baumann RP, Yalamanchili VR, O'Callaghan DJ. Functional mapping and DNA sequence of an equine herpesvirus 1 origin of replication. J Virol 1989 Mar;63(3):1275-83.
- Carter JK, Silva RF. Cell culture amplification of a defective Marek's disease virus. Virus Genes 1990 Sep;4(3):225-37.
- Harty RN, O'Callaghan DJ. An early gene maps within and is 3' coterminal with the immediate-early gene of equine herpesvirus 1. J Virol 1991 Jul;65(7):3829-38.
- Zhao Y, Holden VR, Harty RN, O'Callaghan DJ. Identification and transcriptional analyses of the UL3 and UL4 genes of equine herpesvirus 1, homologs of the ICP27 and glycoprotein K genes of herpes simplex virus. J Virol 1992 Sep;66(9):5363-72.
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