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Registro Completo |
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Biblioteca(s): |
Embrapa Caprinos e Ovinos. |
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Data corrente: |
15/05/2009 |
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Data da última atualização: |
07/12/2023 |
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Autoria: |
TEIXEIRA-GOMES, A. P.; CLOECKAERT, A.; BÉZARD, G.; BOWDEN, R. A.; DUBRAY, G.; ZYGMUNT, M. S. |
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Título: |
Identification and characterization of Brucella ovis immunogenic proteins using two-dimensional electrophoresis and immunoblotting. |
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Ano de publicação: |
1997 |
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Fonte/Imprenta: |
Eletrophoresis, v. 18, p. 1471-1497, 1997. |
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DOI: |
10.1002/elps.1150180824 |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract: In a previous report, proteins from Brucella melitensis were characterized by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and N-terminal microsequencing. In the present report, we have extended this study to the second etiologic agent in ovine brucellosis, B. ovis, responsible for ram epididymitis and infertility. The combination of 2-D gel electrophoresis and protein microsequencing facilitated the location and identification of the major proteins of B. ovis on the 2-D pattern. These proteins comprised cytoplasmic, periplasmic and some membrane proteins except the major outer membrane proteins. By comparing 2-D gel profiles of B. ovis with that of B. melitensis described previously, a few proteins with different expression levels were readily identified. Serum from a ram naturally infected with B. ovis was used in immunoblotting studies to identify immunogenic proteins recognized during the course of infection. This serum showed antibody reactivity against approximately 82 protein spots. Twenty-one of these proteins were identified either by use of monoclonal antibodies or by N-terminal microsequencing. Several proteins previously described in earlier Brucella works were identified: the 89 kDa outer membrane protein, DnaK, GroEL, BP26, and Cu-Zn superoxide dismutase. Eight proteins had amino acid sequences homologous to those of various proteins from other bacteria found in protein databases: NikA, dihydrolipoamide succinyltransferase, a hypothetical 31 kDa protein, malate dehydrogenase, succinyl-CoA synthetase alpha subunit, an amino acid ABC type transporter, Leu/ Ile/ Val-binding protein precursor, and ClpP. The remaining eight proteins had N-terminal sequences lacking similarity to existing database entries. Thus, the 2-D PAGE analysis provided a convenient first approach in the characterization of immunogenic proteins. MenosAbstract: In a previous report, proteins from Brucella melitensis were characterized by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and N-terminal microsequencing. In the present report, we have extended this study to the second etiologic agent in ovine brucellosis, B. ovis, responsible for ram epididymitis and infertility. The combination of 2-D gel electrophoresis and protein microsequencing facilitated the location and identification of the major proteins of B. ovis on the 2-D pattern. These proteins comprised cytoplasmic, periplasmic and some membrane proteins except the major outer membrane proteins. By comparing 2-D gel profiles of B. ovis with that of B. melitensis described previously, a few proteins with different expression levels were readily identified. Serum from a ram naturally infected with B. ovis was used in immunoblotting studies to identify immunogenic proteins recognized during the course of infection. This serum showed antibody reactivity against approximately 82 protein spots. Twenty-one of these proteins were identified either by use of monoclonal antibodies or by N-terminal microsequencing. Several proteins previously described in earlier Brucella works were identified: the 89 kDa outer membrane protein, DnaK, GroEL, BP26, and Cu-Zn superoxide dismutase. Eight proteins had amino acid sequences homologous to those of various proteins from other bacteria found in protein databases: NikA, dihydrolipoamide succinyltransferase, a hypothet... Mostrar Tudo |
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Palavras-Chave: |
Brucella ovis; Brucelosis; Molecular Sequence Data. |
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Thesagro: |
Brucelose. |
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Thesaurus Nal: |
Bacterial antibodies; Bacterial proteins; Brucella melitensis; Immunology; Monoclonal antibodies; Sheep diseases. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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Marc: |
LEADER 02808naa a2200313 a 4500
001 1519568
005 2023-12-07
008 1997 bl uuuu u00u1 u #d
024 7 $a10.1002/elps.1150180824$2DOI
100 1 $aTEIXEIRA-GOMES, A. P.
245 $aIdentification and characterization of Brucella ovis immunogenic proteins using two-dimensional electrophoresis and immunoblotting.
260 $c1997
520 $aAbstract: In a previous report, proteins from Brucella melitensis were characterized by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and N-terminal microsequencing. In the present report, we have extended this study to the second etiologic agent in ovine brucellosis, B. ovis, responsible for ram epididymitis and infertility. The combination of 2-D gel electrophoresis and protein microsequencing facilitated the location and identification of the major proteins of B. ovis on the 2-D pattern. These proteins comprised cytoplasmic, periplasmic and some membrane proteins except the major outer membrane proteins. By comparing 2-D gel profiles of B. ovis with that of B. melitensis described previously, a few proteins with different expression levels were readily identified. Serum from a ram naturally infected with B. ovis was used in immunoblotting studies to identify immunogenic proteins recognized during the course of infection. This serum showed antibody reactivity against approximately 82 protein spots. Twenty-one of these proteins were identified either by use of monoclonal antibodies or by N-terminal microsequencing. Several proteins previously described in earlier Brucella works were identified: the 89 kDa outer membrane protein, DnaK, GroEL, BP26, and Cu-Zn superoxide dismutase. Eight proteins had amino acid sequences homologous to those of various proteins from other bacteria found in protein databases: NikA, dihydrolipoamide succinyltransferase, a hypothetical 31 kDa protein, malate dehydrogenase, succinyl-CoA synthetase alpha subunit, an amino acid ABC type transporter, Leu/ Ile/ Val-binding protein precursor, and ClpP. The remaining eight proteins had N-terminal sequences lacking similarity to existing database entries. Thus, the 2-D PAGE analysis provided a convenient first approach in the characterization of immunogenic proteins.
650 $aBacterial antibodies
650 $aBacterial proteins
650 $aBrucella melitensis
650 $aImmunology
650 $aMonoclonal antibodies
650 $aSheep diseases
650 $aBrucelose
653 $aBrucella ovis
653 $aBrucelosis
653 $aMolecular Sequence Data
700 1 $aCLOECKAERT, A.
700 1 $aBÉZARD, G.
700 1 $aBOWDEN, R. A.
700 1 $aDUBRAY, G.
700 1 $aZYGMUNT, M. S.
773 $tEletrophoresis$gv. 18, p. 1471-1497, 1997.
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Registro original: |
Embrapa Caprinos e Ovinos (CNPC) |
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