Please use this identifier to cite or link to this item: http://www.alice.cnptia.embrapa.br/alice/handle/doc/912252
Research center of Embrapa/Collection: Embrapa Amazônia Ocidental - Resumo em anais de congresso (ALICE)
Date Issued: 2011
Type of Material: Resumo em anais de congresso (ALICE)
Authors: LOBO, I. K. C.
HANADA, R. E.
SOUSA, N. R.
GASPAROTTO, L.
SOUZA Jr. M. T.
KEMA, G. H. J.
SILVA, G. F. da
Additional Information: I. K. C. Lobo; R. E. Hanada, INPA; NELCIMAR REIS SOUSA, CPAA; LUADIR GASPAROTTO, CPAA; EMBRAPA AGROENERGIA; GILVAN FERREIRA DA SILVA, CPAA.
Title: Construction of gateway binary vector for selection with bialaphos or carboxin and GFP expression in fungi.
Publisher: In: CONGRESSO BRASILEIRO DE GENÉTICA, 57., 2011, Águas de Lindóia. Resumos... s.l.: SBG, 2011. p. 25.
Language: pt_BR
Description: 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.
Thesagro: Genética.
Data Created: 2012-01-10
Appears in Collections:Resumo em anais de congresso (CPAA)

Files in This Item:
File Description SizeFormat 
ResumoIgorcoautor.pdf560,65 kBAdobe PDFThumbnail
View/Open

FacebookTwitterDeliciousLinkedInGoogle BookmarksMySpace