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M M Binns

Publications and source records attributed to M M Binns.

At least 91 records · Page 5Linked to original sources
PubMed1990

Insertion of the fusion gene from Newcastle disease virus into a non-essential region in the terminal repeats of fowlpox virus and demonstration of protective immunity induced by the recombinant.

In this paper we report on the identification of non-essential genes in the terminal repeats of the avipox-virus fowlpox virus and the use of these as insertion sites in a vector system. Foreign genes inserted into these sites are shown to be present in two copies in the resultant recombinant virus. To test the potential use of this vector as a live vaccine the fusion gene of Newcastle disease virus has been inserted into a vaccine strain of fowlpox virus and inoculated into chickens. The experiments demonstrate the ability of the recombinant to protect chickens against challenge by a virulent strain of Newcastle disease virus and to elicit the formation of an anti-fusion protein antibody.

Animals↗
PubMed1989

Analysis of the fowlpoxvirus gene encoding the 4b core polypeptide and demonstration that it possesses efficient promoter sequences.

The gene encoding the fowlpox virus 4b core polypeptide has been identified by analogy with the vaccinia 4b gene. It has been cloned, and its nucleotide sequence determined. The gene, which is 1971 nucleotides long, can encode a protein of 75,200 Da (75.2K polypeptide), slightly longer than its vaccinia counterpart with which it shares 52% identity. Sequences upstream of the fowlpox virus 4b gene correspond to the consensus sequence determined for vaccinia late promoters, suggesting that late promoter signals may be shared by the different genera of poxviruses. Upstream sequences have been cloned into a beta-galactosidase translational fusion vector and shown to promote the efficient expression of beta-galactosidase in a transient assay system. This expression was abolished in the presence of araC, an inhibitor of DNA replication which blocks late gene expression in poxviruses. The fowlpoxvirus 4b promoter should be a useful component of genetically engineered fowlpox virus vaccines.

Amino Acid Sequence↗
PubMed1989

Nucleotide sequence of the Marek's disease virus (MDV) RB-1B A antigen gene and the identification of the MDV A antigen as the herpes simplex virus-1 glycoprotein C homologue.

The A antigen gene from a very virulent strain of Marek's disease virus, RB-1B, has been cloned and the nucleotide sequence determined. The predicted amino acid sequence showed 99% identity to that determined for the MDV GA A antigen (Coussens and Velicer, J. Virol. 62, 2373-2379, 1988) over all but the carboxy-terminal region where the sequence diverged extensively. The divergence results from three nucleotide frameshifts in the reported sequence of the MDV GA gene which are not present in a cloned copy of the MDV GA A antigen gene sequenced by us. The MDV A antigen shows significant homology to a number of herpes virus gC homologues, the homology being most extensive in the carboxy-halves of the proteins.

Amino Acid Sequence↗
PubMed1989

Tandem repeated sequences within the terminal region of the fowlpox virus genome.

A 6.2 kb BamHI terminal fragment from fowlpox virus has been cloned and the nucleotide sequence was determined. The fragment was cloned by S1 digestion of viral DNA and therefore does not contain the covalently closed terminal loop. The cloned sequences comprise a short (230 bp) unique region at the terminal end, which is adjacent to a 3.87 bp long, AT-rich region consisting of sets of short tandemly repeated units, 32 and 56 bp long. The remainder of the fragment is composed of a 2.18 kb unique region containing three major open reading frames. The amino acid sequence encoded by one of these has some similarity to that of platelet-derived growth factor.

Amino Acid Sequence↗
PubMed1989

Nucleotide and predicted amino acid sequences of the Marek's disease virus and turkey herpesvirus thymidine kinase genes; comparison with thymidine kinase genes of other herpesviruses.

In this paper we present the nucleotide sequences of the thymidine kinase (TK) genes of two avian herpesviruses: a highly oncogenic strain of Marek's disease virus (MDV strain RB1B) and its serologically related vaccine virus, the herpesvirus of turkeys (HVT strain Fc-126). The predicted coding regions of the two genes are 1029 and 1050 nucleotides respectively, corresponding to polypeptides of 343 and 350 amino acids in length. Putative nucleotide- and nucleoside-binding sites have been identified within the two predicted amino acid sequences. The MDV and HVT TK amino acid sequences exhibit 58.2% amino acid identity. Comparison with other available herpesvirus TK sequences reveals a greater homology to those of the alphaherpesviruses than to those of the gammaherpesviruses. No overall homology was found when compared with the chicken cytoplasmic TK sequence.

Amino Acid Sequence↗
PubMed1989

Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus.

The Marek's disease virus (MDV) homologue of the herpes simplex virus (HSV) gene encoding glycoprotein B (gB) has been identified within BamHI fragments I3 and K3 of the 'highly oncogenic' strain RB1B of MDV. The entire nucleotide sequence of the gene has been determined and its predicted amino acid sequence shown to share gross overall structural features with the gB genes of HSV, varicella-zoster virus (VZV) and other mammalian herpesviruses. In particular, all 10 cysteine residues were conserved in MDV gB and there was extensive homology throughout the gene with VZV, HSV and pseudorabies virus except for the N and C termini. The overall percentage amino acid identity between MDV gB and gB of the alphaherpesviruses had a mean of 50% which was almost twice that between cytomegalovirus and Epstein-Barr virus. Northern blot analysis showed that the main RNA transcribed from this gene is approx. 2.7 kb in size. Antibodies raised against synthetic peptides (residues 250 to 271 and 304 to 330) allowed the identification of a family of serologically related glycoproteins of Mr 110K, 64K and 48K in extracts of MDV-infected cells using immunoblots. Furthermore, the antisera were able to differentiate between the antigens of MDV and herpesvirus of turkeys in immunoblots. Immunofluorescence tests indicated that MDV gB is associated with granules in the cytoplasm and is present at the surface of MDV-infected cells.

Amino Acid Sequence↗
PubMed1988

Non-essential genes in the vaccinia virus HindIII K fragment: a gene related to serine protease inhibitors and a gene related to the 37K vaccinia virus major envelope antigen.

The complete nucleotide sequence of a cloned copy of the HindIII K fragment of the WR strain of vaccinia virus has been determined. Eight open reading frames (ORFs) have been identified, on the basis of size and codon usage. The predicted amino acid sequences of the putative genes have been compared to the Protein Identification Resource and to published vaccinia virus sequences. One gene, predicted to encode a 42.2K protein, is highly related to the family of serine protease inhibitors. It shows approximately 25% identity to human antithrombin III and 19% identity to the cowpox virus 38K protein gene which is also related to serine protease inhibitors. The product of another gene shows a similar high level of identity to the 37K vaccinia virus major envelope antigen. The existence of viable deletion mutants and recombinants containing foreign DNA inserted into both these genes indicates that they are non-essential.

Amino Acid Sequence↗
PubMed1988

Comparison of a conserved region in fowlpox virus and vaccinia virus genomes and the translocation of the fowlpox virus thymidine kinase gene.

The DNA sequence of a clustered set of genes which are conserved in orthopoxviruses has been determined for the avipoxvirus, fowlpox virus. The arrangement of the genes in fowlpox virus is nearly identical to that in vaccinia virus, and genes which are overlapping in vaccinia virus overlap in fowlpox virus. One major difference exists however, as the thymidine kinase (TK) gene is absent in fowlpox virus from the position it occupies within this cluster of genes in vaccinia virus. Instead, in fowlpox virus there is a 32 bp non-coding region present between the genes that flank the TK gene in vaccinia virus. The fowlpox virus TK gene has been cloned and sequenced. The sequences immediately flanking the TK gene show no homology to any previously reported poxvirus gene. These results are discussed in terms of genome stability in poxviruses and the use of the TK gene as a non-essential region for the introduction of foreign genes into poxviruses.

Amino Acid Sequence↗
PubMed1988

Gene sequence and mapping data from Marek's disease virus and herpesvirus of turkeys: implications for herpesvirus classification.

Purified DNAs from Marek's disease virus (MDV) and the herpesvirus of turkeys (HVT) were randomly sheared and cloned into the M13 bacteriophage. Two-hundred and ten MDV and 130 HVT clones were sequenced to give representative samples of the genome sequences. The predicted amino acid sequences from these gammaherpes-viruses were compared to known sequences from other herpesviruses using computer analysis. Thirty-five MDV and 24 HVT genes were identified by comparison with varicella-zoster virus (VZV), an alphaherpesvirus. However, only 14 MDV and seven HVT genes, giving generally lower homology scores, were found by comparison with Epstein-Barr virus (EBV), a gammaherpesvirus, indicating that MDV and HVT sequences bear greater similarity to VZV than to EBV sequences. A number of sequences were mapped by hybridizing labelled M13 clones to Southern blots of restriction fragments of MDV or HVT DNA. The results were consistent with the MDV and HVT genomes being collinear with VZV.

Amino Acid Sequence↗
PubMed1987

An efficient ribosomal frame-shifting signal in the polymerase-encoding region of the coronavirus IBV.

The polymerase-encoding region of the genomic RNA of the coronavirus infectious bronchitis virus (IBV) contains two very large, briefly overlapping open reading frames (ORF), F1 and F2, and it has been suggested on the basis of sequence analysis that expression of the downstream ORF, F2, might be mediated through ribosomal frame-shifting. To examine this possibility a cDNA fragment containing the F1/F2 overlap region was cloned within a marker gene and placed under the control of the bacteriophage SP6 promoter in a recombinant plasmid. Messenger RNA transcribed from this plasmid, when translated in cell-free systems, specified the synthesis of polypeptides whose size was entirely consistent with the products predicted by an efficient ribosomal frame-shifting event within the overlap region. The nature of the products was confirmed by their reactivity with antisera raised against defined portions of the flanking marker gene. This is the first non-retroviral example of ribosomal frame-shifting in higher eukaryotes.

Amino Acid Sequence↗
PubMed1987

Identification by a random sequencing strategy of the fowlpoxvirus DNA polymerase gene, its nucleotide sequence and comparison with other viral DNA polymerases.

The nucleotide sequence of the DNA polymerase gene of the avipoxvirus fowlpox is presented and the predicted amino acid sequence compared with that of the orthopoxvirus vaccinia. The results have brought to light an error in the vaccinia sequence which has resulted in the ommission of 44 amino acids from the carboxy-terminus of the vaccinia DNA polymerase. There has been extensive conservation of amino acids throughout the enzymes, and regions identified as being present in DNA polymerases from a wide range of viruses are again present here. The method used to identify the fowlpoxvirus gene could have applications towards defining genomic organisations in other viral systems.

Amino Acid Sequence↗
PubMed1987

Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus.

The nucleotide sequence determination of the genome of the Beaudette strain of the coronavirus avian infectious bronchitis virus (IBV) has been completed. The complete sequence has been obtained from 17 overlapping cDNA clones, the 5'-most of which contains the leader sequence (as determined by direct sequencing of the genome) and the 3'-most of which contains the poly(A) tail. Approximately 8 kilobases at the 3' end of this sequence have already been published. These contain the sequences of mRNAs A to E within which are the genes for the spike, the membrane and the nucleocapsid polypeptides: the main structural components of the virion. The remainder of the sequence, equivalent to the 'unique' region of mRNA F, is some 20 kilobases in length and is thought to code for a polymerase or polymerases which are involved in the replication of the genome and the production of the subgenomic messenger RNAs. This sequence contains two large open reading frames, potentially coding for polypeptides of molecular weights 441,000 and 300,000. Unlike other large open reading frames in the virus, the 300,000 open reading frame appears to have no subgenomic RNA associated with it which would allow it to be at the 5' end of an mRNA species. Because of this, and because of the characteristics of the sequence in the region immediately upstream of its start codon, other mechanisms of translation, such as ribosome slippage, must be postulated.

Amino Acid Sequence↗
PubMed1987

Expression of the infectious bronchitis virus spike protein by recombinant vaccinia virus and induction of neutralizing antibodies in vaccinated mice.

A cDNA clone of the infectious bronchitis virus (IBV) spike protein gene has been recombined into vaccinia virus. Cells infected with the recombinant virus synthesized IBV spike antigen which was recognized by antibody raised against purified spike protein. Immunofluorescence showed that the IBV spike antigen was transported to the infected cell surface membrane and immunoprecipitation showed the presence of the glycosylated 180K mol. wt. polypeptide precursor of the two spike subunits S1 and S2 that comigrated with this antigen from IBV-infected cells. Vaccinated mice produced antibody that recognized the IBV spike antigen by ELISA and which neutralized IBV infectivity as shown by ciliostasis tests on tracheal organ cultures.

Antibody Formation↗
PubMed1987

Contribution of Salmonella gallinarum large plasmid toward virulence in fowl typhoid.

Four strains of Salmonella gallinarum isolated from independent cases of fowl typhoid all possessed both an 85-kilobase and a 2.5-kilobase plasmid. Each plasmid was eliminated in turn from one of the strains by transposon labeling and curing at 42 degrees C. Elimination of the small plasmid had no effect on the high virulence of the strain for newly hatched and 2-week-old chickens. Whereas oral inoculation of 2-week-old chickens with the parent strain produced 90% mortality with characteristic signs of fowl typhoid, inoculation of the large-plasmid-minus strain produced 0% mortality. A corresponding increase in the 50% lethal dose from log10 1.1 to greater than log10 7.3 was seen with the large-plasmid-minus strain after intramuscular inoculation. Reintroduction of the large plasmid completely restored virulence. A role for the plasmid-linked virulence genes in both invasion and growth in the reticuloendothelial system is suggested by the failure of the large-plasmid-minus strain to penetrate to the liver and spleen after oral inoculation and by its increased clearance from the reticuloendothelial system after intravenous inoculation. These results clearly demonstrate that the large plasmid of S. gallinarum contributes toward virulence in fowl typhoid of chickens.

Animals↗