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 | Acesso ao texto completo restrito à biblioteca da Embrapa Caprinos e Ovinos. Para informações adicionais entre em contato com cnpc.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Caprinos e Ovinos. |
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Data corrente: |
26/02/1996 |
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Data da última atualização: |
01/02/2024 |
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Autoria: |
SEOW, H. F.; ROTHEL, J. S.; PEPIN, M.; DAVID, M. J.; WOOD, P. R. |
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Título: |
Expression, biological activity and kinetcis of production of recombinant ovine TNF-alpha. |
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Ano de publicação: |
1995 |
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Fonte/Imprenta: |
Veterinary Immunology and Immunopathology, v. 44, n. 3/4, p. 279-291, Jan. 1995. |
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DOI: |
10.1016/0165-2427(94)05305-c |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract: Ovine tumour necrosis factor-alpha (OvTNF-α) was cloned by reverse transcription-polymerase reaction using RNA isolated from lipopolysaccharide (LPS)-stimulated alveolar macrophages and primers based on the human TNF-α cDNA sequence. An expression vector carrying the coding sequence of the mature form of ovine TNF was constructed. The recombinant Ov-TNFα (rOvTNF-α) was expressed as a glutathione-S-transferase (GST) fusion protein. It was cleaved with thrombin to yield rOvTNF free of the GST moiety. Growth at a lower temperature of 30°C and use of Escherichia coli strains AM207, AM305, E392 and NM522 did not improve the recovery of rOvTNF-α from the soluble fraction to a significant extent. Purification of recombinant proteins was achieved rapidly and easily by affinity chromatography using glutathione-Sepharose. Yields of pure rOvTNF-α achieved in E. coli JM109 and AM207 were approximately 1 mg L−1. Both rOvTNF-α and recombinant human TNF-α (rhTNF-α) exerted cytotoxicity on L929 cells. However, rOvTNF-α but not rhTNF-α stimulated proliferation of ovine thymocytes. Maximum levels of TNF-α mRNA expression by LPS-stimulated ovine alveolar macrophages were detected at approximately 4 h post-stimulation. |
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Palavras-Chave: |
Base Sequence; Clonagen. |
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Thesagro: |
Biologia Molecular; Doença; Escherichia Coli; Ovino. |
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Thesaurus Nal: |
DNA primers; Gene expression; Genetics; Immunology; Polymerase chain reaction; Recombinant fusion proteins; Sheep diseases; T-lymphocytes. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02249naa a2200349 a 4500
001 1516813
005 2024-02-01
008 1995 bl uuuu u00u1 u #d
024 7 $a10.1016/0165-2427(94)05305-c$2DOI
100 1 $aSEOW, H. F.
245 $aExpression, biological activity and kinetcis of production of recombinant ovine TNF-alpha.$h[electronic resource]
260 $c1995
520 $aAbstract: Ovine tumour necrosis factor-alpha (OvTNF-α) was cloned by reverse transcription-polymerase reaction using RNA isolated from lipopolysaccharide (LPS)-stimulated alveolar macrophages and primers based on the human TNF-α cDNA sequence. An expression vector carrying the coding sequence of the mature form of ovine TNF was constructed. The recombinant Ov-TNFα (rOvTNF-α) was expressed as a glutathione-S-transferase (GST) fusion protein. It was cleaved with thrombin to yield rOvTNF free of the GST moiety. Growth at a lower temperature of 30°C and use of Escherichia coli strains AM207, AM305, E392 and NM522 did not improve the recovery of rOvTNF-α from the soluble fraction to a significant extent. Purification of recombinant proteins was achieved rapidly and easily by affinity chromatography using glutathione-Sepharose. Yields of pure rOvTNF-α achieved in E. coli JM109 and AM207 were approximately 1 mg L−1. Both rOvTNF-α and recombinant human TNF-α (rhTNF-α) exerted cytotoxicity on L929 cells. However, rOvTNF-α but not rhTNF-α stimulated proliferation of ovine thymocytes. Maximum levels of TNF-α mRNA expression by LPS-stimulated ovine alveolar macrophages were detected at approximately 4 h post-stimulation.
650 $aDNA primers
650 $aGene expression
650 $aGenetics
650 $aImmunology
650 $aPolymerase chain reaction
650 $aRecombinant fusion proteins
650 $aSheep diseases
650 $aT-lymphocytes
650 $aBiologia Molecular
650 $aDoença
650 $aEscherichia Coli
650 $aOvino
653 $aBase Sequence
653 $aClonagen
700 1 $aROTHEL, J. S.
700 1 $aPEPIN, M.
700 1 $aDAVID, M. J.
700 1 $aWOOD, P. R.
773 $tVeterinary Immunology and Immunopathology$gv. 44, n. 3/4, p. 279-291, Jan. 1995.
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| 2. |  | PAES, M. C. D.; BARBOSA, N. A.; SILVA, C. S. da; PERUGGIA, E.; PEREIRA FILHO, I. A.; CARLOS, L. de A. Avaliação do milho doce BRSVivi para a produção de conserva de milho verde acidificada. In: CONGRESSO DE PÓS-GRADUAÇÃO DA UFLA, 21., 2012, Lavras. Internacionalização da UFLA: oportunidades e desafios: anais. Lavras: UFLA, 2012.| Tipo: Resumo em Anais de Congresso |
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