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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 107 (1993), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Escherichia coli cultures do not survive the expression of recombinant foot-and-mouth disease virus proteinase 3C. This effect is ascribed to degradation of bacterial protein(s), as concluded from the observation of gradual cessation of gene expression upon induction of 3C expression. Most likely, translation inhibition is the cause of bacterial death, as (i) cell-free translation of the 3C gene was restored by additional bacterial ribosomes, (ii) ribosomes from proteinase 3C-producing cells differed from normal ones by a reduced content of protein S18, and (iii) transcription was not inhibited.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Double-stranded DNA copies of the single-stranded genomic RNA of foot and mouth disease virus have been cloned into the Escherichia coli plasmid pBR322. A restriction map of the viral genome was established and aligned with the biochemical map of foot and mouth disease virus. The coding sequence ...
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  • 3
    ISSN: 1572-994X
    Keywords: expression cloning ; foot-and-mouth disease virus ; non-structural proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Foot-and-mouth disease virus (FMDV) O1 Kaufbeuren-specific cDNA fragments were subcloned into the E. coli expression vector pRIT·2T. Fusion proteins thus produced in bacteria were purified by affinity chromatography and inoculated into rabbits. Three sera thus obtained were found to be monospecific for FMDV proteins 3A, 3C, and 3D, respectively. Two others were prevalently directed against protein 2C, but in addition, either to protein 2B or to protein 3A. Five out of six mature nonstructural virus proteins can therefore be separately investigated in FMDV-infected cells, either by indirect immunofluorescence or by radioimmunoprecipitation. Immunofluorescence shows all investigated proteins to be located exclusively in the cytoplasm. One of them, protein 2C, transiently forms aggregates at the periphery of cells. Radioimmunoprecipitation confirmed current knowledge on maturation of FMDV proteins. It was further used to characterize postinfectional sera with regard to FMDV-specific antibodies. Cattle and guinea pig were found to have responded differently to FMDV nonstructural antigens. Furthermore, antigenicity of yet to be described FMDV polypeptides was observed in the guinea pig.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1572-994X
    Keywords: picornavirus ; foot-and-mouth disease virus ; FMDV ; T7 polymerase ; 3A gene product membrane proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract During picornavirus infection replication of genomic RNA occurs in membrane-associated ribonucleoprotein complexes. These replication complexes contain different nonstructural viral proteins with mostly unknown function. To examine the function of nonstructural picornaviral proteins in more detail, cDNA of foot-and-mouth-disease virus (FMDV) strain O1 Lausanne was cloned into lambda ZAP II, and different parts of the P3-coding sequence were expressed in E. coli by the T7 polymerase system. Expression products constituted (a) fusion proteins composed of N-terminal leader peptide of bacteriophage T7 π10 protein fused to FMDV P3-sequences of different lengths, (b) translation products of authentic P3-region genes, and (c) carboxy-terminally truncated 3A proteins. Expression products were characterized by NaDodSO4-polyacrylamide gel electrophoresis, immunoblotting, as well as electron and immunoelectron microscopy. We show here that in the T7 polymerase system a high level of expression of 3A-containing peptides is achieved in E. coli. Remarkably, the expression of 3A-derived proteins induced a dramatic intracellular membrane proliferation in E. coli cells, similar to the vesicle induction observed in FMDV-infected cells. By immunoelectron microscopy, 3A-reactive material was found associated with these membranes. We hypothesize that the FMDV 3A protein is instrumental in eliciting intracellular membrane proliferation in infected cells as a prerequisite for viral RNA replication.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1572-994X
    Keywords: FMDV subtype O2 ; VP1 ; nucleotide and amino acid sequence ; RNase mismatch cleavage method
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The capsid protein VP1-encoding RNA regions of the foot-and-mouth disease virus isolates O1Lombardy/1946 and O2Brescia/1947 were sequenced and found to be closely related to each other and to O2Normandy/1949, despite some sequence differences. The O1Lombardy sequence was expected to be more closely related to those of the subtype O1 isolates of 1965 and later (e.g., O1Kaufbeuren/1966), but this was not the case. The serological subtyping of both the Lombardy and the Kaufbeuren isolate as O1 strains was possibly due to identical VP1 C-terminal sequences, since all the subtype O2 isolates differ here from the O1 isolates at residue 209. Considerable dissimilarity of other O1Lombardy and O2Brescia genome parts to those of O1Kaufbeuren was qualitatively shown by analyzing the sizes of RNase-treated hybrids formed with virus RNA and defined subgenomic fragments of O1Kaufbeuren-specific antisense cRNA. These hybrids were fragmented into oligonucleotides, but others containing O1Kaufbeuren virus RNA were protected.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Virus genes 16 (1998), S. 185-193 
    ISSN: 1572-994X
    Keywords: foot-and-mouth disease virus ; VP1 ; sequence ; RT-PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A large part of the capsid protein VP1-coding sequence of foot-and-mouth disease virus, isolated between 1993 and 1996 in Europe, was amplified by the reverse transcription-dependent polymerase chain reaction (RT-PCR). The same was done with some non-European virus isolates, especially those against which vaccines were currently produced. The products were sequenced, and the sequences aligned. The alignment comprizes sequences of the types A, O and Asia1. Although the provenance of virus introduced to Europe remains unknown, genetic relation to some other isolates was indicated. Several genotypes of the virus were found to circulate in the field since years.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1572-994X
    Keywords: type A and O FMDV ; VP1-coding sequence ; monoclonal antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This report extends the knowledge on the epizootical situation of foot-and-mouth disease in Asia. RNA from six samples of type A and five of type O virus, isolated between 1987 and 1997 in Bangladesh, Iran, Malaysia and Turkey, was subjected to reverse transcription-dependent polymerase chain reactions that amplify large parts of the capsid protein VP1 encoding genome region. The amplification products were sequenced, and the sequences aligned to each other and to published sequences. This showed the type O isolates of 1987–1997 from Bangladesh to be of same genotype and closely related to isolates of 1988 and later from Saudi Arabia, 1990 from India, 1996 from Greece and Bulgaria, and 1997 from Iran. Among the analyzed type A isolates, those of 1992 and 1996 from Turkey were of same genotype and related to previously described isolates of 1987 from Iran and of 1992 from Saudi Arabia. The isolate of 1997 from Malaysia was found to be related to isolates from Thailand of 1993 and 1996. The isolates of 1987 from Bangladesh and 1997 from Iran, however, represent different so far not described genotypes. Monoclonal antibodies, raised against the vaccine production strains A22 Iraq, Asia1 Shamir, O1 Kaufbeuren and O1 Manisa, and the recent type A field isolates Saudi Arabia/92 and Albania/96, were used in an ELISA to compare the reaction patterns of many of the field isolates. The monoclonal antibodies were further characterized for virus-neutralizing activity and binding to trypsinized homologous virus. The failure of neutralizing antibodies in binding to trypsinized homologous as well as to heterologous virus suggested the epitopes to reside at the major antigenic component of the virus, which is the capsid protein VP1. Two non-neutralizing antibodies that bind to trypsin-sensitive epitopes cross-reacted, however, with heterologous virus. This indicates the existence of a trypsin-sensitive antigenic site outside of VP1. In summary, the results obtained by ELISA confirm the observed sequence differences, but indicate further sequence differences at minor antigenic sites that do not reside on VP1.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Virus genes 2 (1989), S. 283-291 
    ISSN: 1572-994X
    Keywords: FMDV A5Bernbeuren/1984 ; FMDV A Iran/1987 ; VP1 ; sequences ; 2-D structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have sequenced the nucleotides of the regions that encode the capsid protein VP1 of the foot-and-mouth disease viruses (FMDV) A5Bernbeuren/1984 and A Iran/1987. Amino acid sequences and secondary protein structures are provided. Both proteins consist of 212 amino acids. The sequences and secondary structures are compared to those of FMDV A22/CCCP/64, a strain previously endemic in the Near East. Nucleotide divergency among the three sequences is highest for FMDV A5Bernbeuren/1984 (18% compared to 13% for each other case). Thirty amino acid divergencies are observed between A22/CCCP/64 and A5 Bernbeuren/1984 or A Iran/1987, whereas the latter two differ by 27 residues. The secondary structures of all three proteins are different. A Iran/1987 is considered to belong to a thus far unknown subtype.
    Type of Medium: Electronic Resource
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