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Title: Optimization of RQRT-PCR protocols to measure -1,3-glucanase mRNA levels in infected tissues of rubber tree (Hevea brasiliensis)
Authors: Thanseem, I
Thulaseedharan, A
Keywords: Actin
Hevea brasiliensis
Relative RT-PCR
Issue Date: Jun-2006
Publisher: CSIR
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 -1,3-glucanase gene was studied in the pathogen infected tissues of rubber. The -1,3-glucanase gene was found to be induced in infected leaf tissues and reached a peak at 48 h after inoculation. The -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.
Page(s): 492-498
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Source:IJEB Vol.44(06) [June 2006]

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