Restriction Endonuclease Analysis of Bovine Herpesvirus 1 DNA and Nucleic Acid Homology between Isolates

Abstract

SUMMARYIsolates of bovine herpesvirus 1 (BHV-1) are associated with a variety of clinical manifestations. To determine if a single form of BHV-I was responsible for the different virus-associated diseases or whether subpopulations of various isolates produced different clinical symptoms, studies were initiated to examine the DNA restriction enzyme patterns and nucleic acid homology between virus isolates from respiratory infections and other clinical syndromes. Differences between the genomes of several virus isolates were detected using DNA restriction enzyme analyses. However, nucleic acid hybridization studies of the virus DNAs using filter and liquid hybridization indicated at least a 95~ genetic homology between the virus isolates from different types of infections. Additionally, these studies demonstrated that the DNA of BHV-1 had an average molecular weight of 84 x 106.Bovine herpesvirus 1 (BHV-1) has been associated with a variety of clinical syndromes. These are primarily respiratory, but may also inclfide genital infections, encephalitis, ocular carcinoma and abortion. The virus associated with respiratory diseases has been referred to as infectious bovine rhinotracheitis (IBR) virus, whereas the virus isolate associated with genital infections has been termed infectious pustular vulvovaginitis (IPV) virus (Kahrs, 1977;Pastoret et al., 1982). Misra et al. (1983) reported that isolates of BHV-I could be divided into three distinct groups based upon the virus DNA restriction enzyme patterns. As in the situation of herpes simplex virus types 1 and 2 in humans, the possibility exists that respiratory and genital infections may be caused by partially related viruses that appear serologically similar, but by analysis of the virus DNA represent different virus types. Consequently, the investigations reported here were initiated to determine the amount of genetic homology between a respiratory isolate of BHV-1 and other clinical isolates using restriction enzyme analysis and nucleic acid hybridization of virus DNA.Since reports indicated that DNA from BHV-1 isolates associated with respiratory infections (IBR virus) had different restriction patterns from the DNA isolated from viruses associated with genital infections (IPV virus) (Engels et al., 1981), we initially differentiated virus isolates using restriction enzyme analysis and nucleic acid hybridization. The BHV-1 isolates used in these studies were IBR virus Los Angeles (LA), a respiratory disease isolate, obtained from the American Type Culture Collection and an IPV virus isolate obtained from the Department of Microbiology, School of Veterinary Medicine, University of California, Davis. A variety of isolates from various clinical disease states were also obtained from the School of Veterinary Medicine at Washington State University, Pullman, Washington and from the Agriculture Experiment Station at the University of Nevada. Bovine embryonic lung cells were used for all experiments and cultured in Dulbecco's MEM with 5 ~ foetal calf serum. Virus DNA fo...

The location and nucleotide sequence of the thymidine kinase gene of bovine herpesvirus type 1.2

“…Restriction endonuclease (RE) profiles and clinical manifestations have been used to classify BHV-1 strains into IBR-like and IPV-like profiles (Engels et al, 1981 ;Seal et al, 1985;Osorio et al, 1985). Although it provides a useful method of sub-classification, no significant biological difference can be attributed to BHV-1 strains isolated from the respiratory and genital tracts.…”

Restriction Endonuclease Analysis of Alcelaphine Herpesvirus 1 DNA and Molecular Cloning of Virus Genomic DNA for Potential Diagnostic Use

“…39 The Los Angeles isolate of bovine herpesvirus 1 (BHV-1) has been extensively characterized. 21,43 Bovine herpesvirus 2(BHV-2) a4 and the Movar isolate of bovine herpesvirus 4(BHV-4) 8,13,47 have been previously described.…”

“…Gels were stained with ethidium bromide and photographed with Polaroid type 55 film during UV transillumination. 43 For blot hybridization analysis, DNA fragments were transferred to 0.45-µm pore size nitrocellulose 46 and hybridized in 50% formamide at 37 C 19 with nick-translated 32 AHV-1 isolate DNA or the designated cloned DNA probe as previously described. 43 Nitrocellulose filters f with blotted virus DNA fragments were hybridized with 32 P-labeled probe DNA.…”

Genomic Heterogeneities in Bovine Herpesvirus Type 1 Viral Isolates: A Major Variant Selected from a Field Isolate

“…By restriction en donuclease analysis, only one fragment (the EcoR\ fragment C) is found to be heteroge neous in size within the strain I isolates ex amined [7]. Furthermore, several authors have used restriction endonuclease analysis to compare various isolates of BHV-1 [5,6,[8][9][10][14][15][16][17], and no difference was noted between the IBR isolates. However, the res triction enzymes used in these studies were mainly //mdIII, BamHl, Hpal, and £coRI, for which there are very few restriction sites (12,8,6, and 6, respectively) [13] in the IBR genome.…”

“…In an effort to differentiate BHV-I clinical isolates, their polypeptide profiles, antigenic characteristics, and genomic restriction en donuclease patterns have been compared [2,[4][5][6][7][8][9][10]. It generally is accepted that BHV-1 isolates are divided into three types/subtypes [6,9].…”