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M M Binns

Publications and source records attributed to M M Binns.

At least 19 recordsLinked to original sources
PubMed1996

Antigenic and genetic evolution of equine H3N8 influenza A viruses.

Evolution of equine influenza a H3N8 viruses was examined by antigenic and genetic analysis of a collection isolates from around the world. It was noted that antigenic and genetic variants of equine H3N8 viruses cocirculate, and in particular that variants currently circulating in Europe and the USA are distinguishable from one another both in terms of antigenic reactivity and genetic structure of the HA1 portion of the haemagglutinin (HA) molecule. Whilst the divergent evolution of American and European isolates may be due to geographical isolation of the two gene pools, some mixing is believed to occur as 'American-like' viruses have been isolated during outbreaks of equine influenza in the UK. The cocirculation of two antigenically and genetically distinct lineages of equine influenza H3N8 viruses has serious implications for vaccine strain selection.

Amino Acid Sequence↗
PubMed1995

Development of PCR assays to detect genetic variation amongst equine herpesvirus-1 isolates as an aid to epidemiological investigation.

A search for variable restriction sites has been carried out for equine herpesvirus-1 (EHV-1) in an attempt to develop markers which can be used to group epidemiologically related viruses into groups, and to learn more about the dynamics of EHV-1 disease. Crude viral DNA extracts of EHV-1, prepared by Hirt extraction, were digested with AluI, HaeIII, or RsaI, and Southern blotted following electrophoresis. DNA fingerprints, produced by probing the Southern blots with the EHV-1 EcoR1-I fragment, separated 56 isolates into 16 groups. The variable sites within the EcoR1-I fragment were mapped approximately using fragments from within EcoR1-I, and the precise location of the variable sites determined from the DNA sequence of this fragment. Oligonucleotide primers flanking the variable sites were synthesized, and used in PCR assays to detect variable fragments. The AluI variable fragment was found to result from the presence or absence of a single AluI site. In contrast, the variable bands seen with HaeIII and RsaI, resulted from variation in the copy number of two tandemly repeated sequences, one of which had not previously been recognized. In addition, HaeIII digests of EHV-1 isolates probed with the glycoprotein B (gB) gene of EHV-1 also separated isolates into two groups. The variable HaeIII site was mapped towards the 5'-end of the gB gene and a PCR assay established. The distribution of the variable AluI site within the EcoR1-I fragment and the HaeIII site within the gB gene were estimated on a large number of clinical isolates using PCR on unpurified viral tissue culture medium. Both sites had a good distribution and together with additional variable sites should provide the basis for the rapid DNA fingerprinting of EHV-1 isolates.

Animals↗
PubMed1995

Replication of equid herpesvirus 4 in endothelial cells and synovia of a field case of viral pneumonia and synovitis in a foal.

Equid herpesvirus 4 (EHV-4) infection was diagnosed as the cause of interstitial pneumonia in a 6-week-old conventionally reared Welsh pony foal, by cocultivation and immunolabelling with specific monoclonal antibodies, EHV-4 specific amplification of viral DNA, and immunohistological examination of infected tissues. The case was novel in that replication of the EHV-4 isolate in endothelial cells and in the synovial epithelium was a feature. Restriction digests of this isolate were compared with those of seven respiratory and one abortigenic EHV-4 isolate, and no differences in restriction pattern were evident. The implications of these findings for the pathogenesis of EHV-4 infection are discussed.

Animals↗
PubMed1995

Dog parentage testing using canine microsatellites.

Canine microsatellites, comprising one to four base pair repeated sequences, were identified as part of a project to generate a genetic linkage map of the dog. They have been used to assign parentage in a range of doubtful paternity cases, for example, all the pups in a litter from a bitch mated on different days to two different stud dogs were identified by DNA analysis to come from one dog only. DNA typing in another case was able to exclude a male dog as the father of a litter. DNA analysis can be of relevance by allowing pedigree dogs, which would have previously been excluded on the basis of unknown paternity, to be registered with the appropriate kennel club.

Animals↗
PubMed1994

Comparison of M and N gene sequences distinguishes variation amongst equine arteritis virus isolates.

cDNA copies of the M and N genes of equine arteritis virus (EAV) isolates were synthesized by reverse transcription followed by polymerase chain reaction amplification. The cDNA was subjected to a cycle sequencing strategy using Taq polymerase, and the nucleotide and derived amino acid sequences of 10 virus isolates were compared. The M and N genes of all isolates had the same initiation and termination sites as the prototype Bucyrus strain and the encoded proteins were conserved between viruses. Comparison of nucleotide sequence homologies and phylogenetic tree analysis implied the existence of three EAV variants originating from the U.S.A. (Bucyrus), Austria (Vienna) and Switzerland (Bibuna), and suggested that RNA recombination between EAV isolates may have occurred.

Base Sequence↗
PubMed1994

Deletion of fowlpox virus homologues of vaccinia virus genes between the 3 beta-hydroxysteroid dehydrogenase (A44L) and DNA ligase (A50R) genes.

A fragment of 4156 bp of fowlpox virus (FPV) genomic DNA contains homologues of vaccinia virus 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta-HSD; A44L) and DNA ligase (A50R) genes. The FPV locus has clearly been rearranged relative to that of vaccinia virus as homologues of genes A45R to A49R, including the thymidylate kinase and a gene with homology to superoxide dismutase, are deleted. The deleted genes are replaced by two open reading frames: for a serine proteinase inhibitor with homology to vaccinia virus gene K2L and for a protein with no significant homology to proteins in the databases. In addition, the FPV homologues of A44L and A50R are in the same polarity in FPV whereas they are in opposite polarities in vaccinia virus. Increased 3 beta-HSD activity has been demonstrated in cells infected with either of two different strains of FPV or with canarypox virus.

3-Hydroxysteroid Dehydrogenases↗
PubMed1993

Characterization of canine microsatellites.

Canine DNA was cloned in M13 and screened for the presence of (dC-dA)n.(dG-dT)n repeats. Oligonucleotide primers were synthesised to the microsatellite flanking sequences and used in the polymerase chain reaction to amplify those loci from genomic DNA. The polymorphism of each microsatellite was estimated in a set of unrelated dogs.

Animals↗
PubMed1993

Genetic and antigenic analysis of an equine influenza H 3 isolate from the 1989 epidemic.

The haemagglutinin (HA) gene from the equine influenza H3N8 isolate Suffolk/89 has been cloned by reverse transcription and polymerase chain reaction amplification. The nucleotide sequence of the HA gene was determined from two independently cloned copies of the gene and was found to be most closely related to recent American isolates supporting the idea that most isolates of equine H3N8 are evolving as a single lineage. When the predicted amino acid sequence of the Suffolk/89 HA was examined, changes had taken place in at least four of the major antigenic sites, A, B, C, and D when compared to the sequences of the isolates used in the current vaccines (Miami/63 and Fontainebleau/79). Surprisingly, when the Suffolk/89 isolate was tested in haemagglutination inhibition (HI) assays with a panel of six mouse monoclonal antibodies, no differences were observed between the Suffolk/89 and the Fontainebleau/79 isolates, suggesting that this panel of monoclonal antibodies may recognise a limited subset of the major antigenic sites. Three anti-HA horse heterohybridoma monoclonals were able to distinguish between the Suffolk/89 and Fontainebleau/79 viruses, demonstrating that the horse does recognise these isolates as being antigenically different. The results of the work suggest that the isolates used in current equine influenza vaccines may need updating.

Amino Acid Sequence↗
PubMed1993

Alterations in DNA sequence and RNA transcription of the Bam HI-H fragment accompany attenuation of oncogenic Marek's disease herpesvirus.

The nucleotide sequences of the BamHI-H fragment of the HPRS16 strain of Marek's disease virus (MDV) and of its attenuated derivative HPRS16/att have been determined. The results show that in addition to the tandem expansion of a 132 bp sequence from two copies in HPRS16 virus to eight copies in HPRS16/att, nucleotide substitutions, deletions, and insertions were also noted. Several potential open reading frames (ORFs) were identified. One of these (ORF 13) encoded a deduced protein, mol wt 32 kD, which is likely to be the serotype-1 specific phosphoprotein expressed in tumours (1) and mapped to an EcoRI fragment within BamHI-H (2). Our results suggest that this ORF is unlikely to be the B antigen (3). ORF 4, which had some similarity to CD4 and immunoglobulin M heavy chain, was encoded by a transcript that originated within the first copy of the 132 bp repeat. ORF 21, which mapped entirely within UL, encoded a deduced protein at least 322 amino acids long that had some similarity to varicella zoster virus (VZV) alpha trans-inducing protein. None of these ORFs was altered significantly by attenuation, except ORF 4 and another small ORF (ORF 3), 5' of the 132 bp repeats, which would probably fail to be transcribed because of truncation of an RNA transcript.

Amino Acid Sequence↗
PubMed1993

Construction and properties of a turkey herpesvirus recombinant expressing the Marek's disease virus homologue of glycoprotein B of herpes simplex virus.

A herpesvirus of turkeys (HVT) recombinant containing a 3.9 kbp fragment of Marek's disease virus (MDV) DNA encoding MDV glycoprotein B (gB), stably integrated into the thymidine kinase (TK) gene of HVT, has been constructed. The replication of the recombinant in chick embryo fibroblasts (CEF) was comparable to that of wild-type HVT. The recombinant expressed authentic MDV gB and its processed forms (110K, 65K and 48K) in CEF as shown by immunoblotting using an MDV-specific anti-peptide serum. Northern blot analysis showed that MDV gB mRNA was transcribed from MDV promoter sequences flanking the MDV gB open reading frame and also from the HVT TK promoter. However, the level of replication of the recombinant in vivo appeared to be lower than wild-type HVT as shown by the titres of HVT antibodies, determined by ELISA. Pathogenicity tests showed that the recombinant was safe and did not cause microscopic or gross Marek's disease lesions or other abnormalities. The results suggest that HVT has potential as a vector for recombinant vaccines.

Animals↗
PubMed1993

Identification and sequence analysis of the homologues of the herpes simplex virus type 1 glycoprotein H in Marek's disease virus and the herpesvirus of turkeys.

The glycoprotein H (gH) genes of two avian herpesviruses, Marek's disease virus and the herpesvirus of turkeys, have been cloned and sequenced and the coding regions found to be of 2439 and 2424 nucleotides respectively. The predicted primary polypeptide products of these open reading frames are 813 and 808 amino acids and correspond to Mrs of 90,800 and 91,100. Both amino acid sequences exhibit characteristic glycoprotein features such as hydrophobic signal and anchor sequences and potential sites for N-linked glycosylation. Polypeptide sequence comparison to the other eight available gH sequences revealed more similarity to the alphaherpesvirus subgroup than to either beta- or gammaherpesviruses.

Amino Acid Sequence↗
PubMed1993

Isolation and characterization of microsatellites from the canine genome.

Microsatellite sequences comprising (dC-dA)n.(dG-dT)n repeats have been isolated from canine libraries and sequenced. Oligonucleotide primers have been synthesized to the microsatellite flanking sequences and used in the polymerase chain reaction to amplify those loci from genomic DNA. The degree of polymorphism of each microsatellite was estimated in a set of unrelated dogs. It is concluded that of the 10 loci studied, nine are sufficiently polymorphic to be useful in genetic studies.

Animals↗
PubMed1993

Characterisation of equine influenza isolates from the 1987 epizootic in India by nucleotide sequencing of the HA1 gene.

Two A/Equi-2 (H3N8) isolates were obtained during the 1987 Indian equine influenza epizootic. The sequence of the Ludhiana/87 HA1 gene revealed that this isolate was very similar to recent European and North American isolates of equine influenza. In contrast, the Bhiwani/87 HA1 gene was nearly identical to the Miami/63 prototype H3 sequence. These results support the antigenic analysis previously carried out on these isolates using monoclonal antibodies. However, the finding that Bhiwani/87 is so similar to Miami/63, coupled with the finding that equine H3N8 influenza viruses have previously been shown to evolve along a single lineage, suggests that Miami/63 virus from either a vaccine or laboratory source may be the origin of the Bhiwani/87 isolate.

Amino Acid Sequence↗
PubMed1993

Detection of antibodies against equine herpesvirus types 1 and 4 by using recombinant protein derived from an immunodominant region of glycoprotein B.

The N-terminal fragment comprising residues +1 to +50 (gB1-50) of equine herpesvirus type 1 (EHV-1) glycoprotein B was expressed as a glutathione S-transferase fusion protein in Escherichia coli. Recombinant gB1-50 (rgB1-50) was recognized in immunoblots by sera from rabbits immunized with EHV-1 and by convalescent-phase sera from horses with natural EHV-1 infections. An enzyme-linked immunosorbent assay (ELISA) for monitoring antibody levels against EHV-1 was developed by using rgB1-50, and its specificity was assessed with a panel of reference antisera against other equine viruses. A specific cross-reaction was detected with EHV-4, which was confirmed by inhibition ELISA. Convalescent-phase sera from horses with natural EHV-1 or EHV-4 infections possessed antibody titers against rgB1-50 ranging from 1:2,000 to 1:64,000, indicating the presence of an immunodominant antigenic site. The study demonstrated the potential application of rgB1-50 as a diagnostic antigen and highlights the glutathione S-transferase fusion system as a simple and effective method of producing purified milligram quantities of antigen.

Amino Acid Sequence↗