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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
28/08/2018 |
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Data da última atualização: |
12/09/2018 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
TOSCANO, J. H. B.; LOPES, L. G.; GIRALDELO, L. A.; SILVA, M. H. da; OKINO, C. H.; CHAGAS, A. C. de S. |
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Afiliação: |
João Henrique Barbosa Toscano, UNESP; Louyse Gabrielli Lopes, UNICEP; Luciana Aparecida Giraldelo, UNICEP; Matheus Henrique da Silva, UNICEP; CINTIA HIROMI OKINO, CPPSE; ANA CAROLINA DE SOUZA CHAGAS, CPPSE. |
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Título: |
Identification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus. |
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Ano de publicação: |
2018 |
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Fonte/Imprenta: |
Molecular Biology Reports, v.45, n.5, p.1253-1262, out. 2018. |
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Idioma: |
Inglês |
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Conteúdo: |
Due to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results. MenosDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes,... Mostrar Tudo |
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Palavras-Chave: |
Genes de referência. |
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Thesagro: |
Haemonchus Contortus; Ovino. |
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Thesaurus Nal: |
Sheep. |
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Categoria do assunto: |
G Melhoramento Genético |
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Marc: |
LEADER 02463naa a2200229 a 4500
001 2094611
005 2018-09-12
008 2018 bl uuuu u00u1 u #d
100 1 $aTOSCANO, J. H. B.
245 $aIdentification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus.$h[electronic resource]
260 $c2018
520 $aDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results.
650 $aSheep
650 $aHaemonchus Contortus
650 $aOvino
653 $aGenes de referência
700 1 $aLOPES, L. G.
700 1 $aGIRALDELO, L. A.
700 1 $aSILVA, M. H. da
700 1 $aOKINO, C. H.
700 1 $aCHAGAS, A. C. de S.
773 $tMolecular Biology Reports$gv.45, n.5, p.1253-1262, out. 2018.
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| 3. |  | POLICARPO, V. M.; ENDO, E.; FARIA, R. Q.; ANJOS, V.; BORGES, S. V.; GREGÓRIO, S. R.; CAVALCANTI, N. de B. Efecto de aditivos sobre el calor, textura y aceptación del dulce de umbu (Spondias tuberosa, Arr.Cam.) verde. Alimentaria, Madrid, n. 346, p. 111-116, 2003.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: Internacional - B |
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