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Registro Completo |
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Biblioteca(s): |
Embrapa Amazônia Ocidental. |
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Data corrente: |
10/01/2012 |
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Data da última atualização: |
22/04/2021 |
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Tipo da produção científica: |
Resumo em Anais de Congresso |
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Autoria: |
LOBO, I. K. C.; HANADA, R. E.; SOUSA, N. R.; GASPAROTTO, L.; SOUZA Jr. M. T.; KEMA, G. H. J.; SILVA, G. F. da. |
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Afiliação: |
I. K. C. Lobo; R. E. Hanada, INPA; NELCIMAR REIS SOUSA, CPAA; LUADIR GASPAROTTO, CPAA; EMBRAPA AGROENERGIA; GILVAN FERREIRA DA SILVA, CPAA. |
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Título: |
Construction of gateway binary vector for selection with bialaphos or carboxin and GFP expression in fungi. |
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Ano de publicação: |
2011 |
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Fonte/Imprenta: |
In: CONGRESSO BRASILEIRO DE GENÉTICA, 57., 2011, Águas de Lindóia. Resumos... s.l.: SBG, 2011. p. 25. |
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Idioma: |
Português |
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Conteúdo: |
Genomic data has created a growing demand for tools and methodologies for studying the genes function, which can be realized through loss of function experiments (gene knockout) or by RNA silencing (knockdown). The develop-ment of binary vectors for Agrobacterium tumefaciens mediated transformation (ATMT) has the advantage of being independent of protoplast formation and can be used directly on a wide variety of fungal species and tissue types. The selection of transformants using bialaphos and carboxin has the advantages of low cost in the transformation and availability of different selectable markers, also allowing the analysis of several genes and combination of study by knockout or knockdown, using selectable markers in the same transformant. Thus, this study aimed to build two binary vectors containing reporter gene and selectable markers that confer resistance to carboxin and bialaphos. The cassettes were constructed using the Gateway system to two fragments. The gene encoding the GFP protein and PtoxA and PtrpCpromoters were cloned into pDONR P1-P5R plasmid. Genes that confer bialaphos and carboxin resistance, bar and cbxr respectively, were cloned into pDONR P5-P2 plasmid. The pPGW plasmid was used as des-tination vector. The gfp gene transcription is controlled by PtoxA promoter and the bar and cbxr genes transcriptions are controlled by PtrpC promoter. These binary vectors were named pGWGFP-BAR and pGWGFP-CBXR. The assembly of cassettes was confi rmed by sequencing, and the validation of vectors is being accomplished through transformation (ATMT) with the plant pathogens Mycosphaerella fi jiensis and Fusarium oxysporum f. sp. cubense. MenosGenomic data has created a growing demand for tools and methodologies for studying the genes function, which can be realized through loss of function experiments (gene knockout) or by RNA silencing (knockdown). The develop-ment of binary vectors for Agrobacterium tumefaciens mediated transformation (ATMT) has the advantage of being independent of protoplast formation and can be used directly on a wide variety of fungal species and tissue types. The selection of transformants using bialaphos and carboxin has the advantages of low cost in the transformation and availability of different selectable markers, also allowing the analysis of several genes and combination of study by knockout or knockdown, using selectable markers in the same transformant. Thus, this study aimed to build two binary vectors containing reporter gene and selectable markers that confer resistance to carboxin and bialaphos. The cassettes were constructed using the Gateway system to two fragments. The gene encoding the GFP protein and PtoxA and PtrpCpromoters were cloned into pDONR P1-P5R plasmid. Genes that confer bialaphos and carboxin resistance, bar and cbxr respectively, were cloned into pDONR P5-P2 plasmid. The pPGW plasmid was used as des-tination vector. The gfp gene transcription is controlled by PtoxA promoter and the bar and cbxr genes transcriptions are controlled by PtrpC promoter. These binary vectors were named pGWGFP-BAR and pGWGFP-CBXR. The assembly of cassettes was confi rmed by sequencin... Mostrar Tudo |
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Thesagro: |
Genética. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/51989/1/Resumo-Igor-co-autor.pdf
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Marc: |
LEADER 02305nam a2200193 a 4500
001 1912252
005 2021-04-22
008 2011 bl uuuu u00u1 u #d
100 1 $aLOBO, I. K. C.
245 $aConstruction of gateway binary vector for selection with bialaphos or carboxin and GFP expression in fungi.
260 $aIn: CONGRESSO BRASILEIRO DE GENÉTICA, 57., 2011, Águas de Lindóia. Resumos... s.l.: SBG, 2011. p. 25.$c2011
520 $aGenomic data has created a growing demand for tools and methodologies for studying the genes function, which can be realized through loss of function experiments (gene knockout) or by RNA silencing (knockdown). The develop-ment of binary vectors for Agrobacterium tumefaciens mediated transformation (ATMT) has the advantage of being independent of protoplast formation and can be used directly on a wide variety of fungal species and tissue types. The selection of transformants using bialaphos and carboxin has the advantages of low cost in the transformation and availability of different selectable markers, also allowing the analysis of several genes and combination of study by knockout or knockdown, using selectable markers in the same transformant. Thus, this study aimed to build two binary vectors containing reporter gene and selectable markers that confer resistance to carboxin and bialaphos. The cassettes were constructed using the Gateway system to two fragments. The gene encoding the GFP protein and PtoxA and PtrpCpromoters were cloned into pDONR P1-P5R plasmid. Genes that confer bialaphos and carboxin resistance, bar and cbxr respectively, were cloned into pDONR P5-P2 plasmid. The pPGW plasmid was used as des-tination vector. The gfp gene transcription is controlled by PtoxA promoter and the bar and cbxr genes transcriptions are controlled by PtrpC promoter. These binary vectors were named pGWGFP-BAR and pGWGFP-CBXR. The assembly of cassettes was confi rmed by sequencing, and the validation of vectors is being accomplished through transformation (ATMT) with the plant pathogens Mycosphaerella fi jiensis and Fusarium oxysporum f. sp. cubense.
650 $aGenética
700 1 $aHANADA, R. E.
700 1 $aSOUSA, N. R.
700 1 $aGASPAROTTO, L.
700 1 $aSOUZA Jr. M. T.
700 1 $aKEMA, G. H. J.
700 1 $aSILVA, G. F. da
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Registro original: |
Embrapa Amazônia Ocidental (CPAA) |
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| 1. |  | LOBO, I. K. C.; HANADA, R. E.; SOUSA, N. R.; GASPAROTTO, L.; SOUZA Jr. M. T.; KEMA, G. H. J.; SILVA, G. F. da. Construction of gateway binary vector for selection with bialaphos or carboxin and GFP expression in fungi. In: CONGRESSO BRASILEIRO DE GENÉTICA, 57., 2011, Águas de Lindóia. Resumos... s.l.: SBG, 2011. p. 25.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Ocidental. |
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