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Optimization of RQRT-pCR protocols to measure beta-1,3-glucanase mRNA levels in infected tissues of rubber tree (Hevea brasiliensis).
Indian J Exp Biol ; 2006 Jun; 44(6): 492-8
Artigo em Inglês | IMSEAR | ID: sea-55813
ABSTRACT
RQRT-PCR technique was evaluated for its validity as an alternative to Northern blotting for quantification of plant gene expression in diseased tissues of Hevea. Reliable RT-PCR results could be obtained by co-amplification of housekeeping actin gene as the internal control along with the gene of interest. The product of interest was quantified relative to that of the internal control by measuring net intensity of bands. Expression levels of defense-related beta-1,3-glucanase gene was studied in the pathogen infected tissues of rubber. The beta-1,3-glucanase gene was found to be induced in infected leaf tissues and reached a peak at 48 h after inoculation. The beta-1,3-glucanase gene expression during pathogen infection was determined through Northern blot hybridization also, using 18S RNA as the internal control. RQRT-PCR and Northern hybridization showed almost similar results, thereby validating the use of this technique to study the gene expression in rubber.
Assuntos
Texto completo: DisponíveL Índice: IMSEAR (Sudeste Asiático) Assunto principal: Phytophthora / Borracha / Autorradiografia / Fatores de Tempo / RNA / RNA Mensageiro / Northern Blotting / Southern Blotting / DNA Complementar / Folhas de Planta País/Região como assunto: América do Sul / Brasil Idioma: Inglês Revista: Indian J Exp Biol Ano de publicação: 2006 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: IMSEAR (Sudeste Asiático) Assunto principal: Phytophthora / Borracha / Autorradiografia / Fatores de Tempo / RNA / RNA Mensageiro / Northern Blotting / Southern Blotting / DNA Complementar / Folhas de Planta País/Região como assunto: América do Sul / Brasil Idioma: Inglês Revista: Indian J Exp Biol Ano de publicação: 2006 Tipo de documento: Artigo