Barsalobres-Cavallari, C.F. , Maluf, M.P. , Maia, I.G. .
Validating internal controls for gene expression studies in Coffea arabica.
COFFEE belongs to the genus Coffea in the family Rubiaceae, and is mostly grown in tropical and subtropical regions. About 100 species of the genus Coffea have been identified so far, but commercial production relies only on two species, Coffea arabica and Coffea canephora. C. arabica is the only allopolyploid (2n = 4x = 44) coffee species and self-fertile. The other Coffea species are diploid (2n = 2x = 22) and generally self-sterile. In this study, ten housekeeping genes representing different functional classes and gene families were chosen for investigation of gene expression using quantitative reverse transcription PCR (RT-PCR) analysis. These include ubiquitins, actins, tubulins, translational initiation factor, rRNA, and others. Searches in the literature revealed that members of all classes have been used as internal controls for studies of plant gene expression by RNA gel blots or RT-PCR assays. The former set of coffee genes was identified in silico using the Brazilian Coffee EST database. The expression patterns of these genes were determined in five tissue samples. Analysis showed that the potential internal control genes differed significantly in their expression levels on the tested tissues. Our results support that quantitative comparisons of candidate reference genes are an important part of RT-PCR studies that seek to precisely evaluate variation in gene expression.
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1 - UNESP - Campus de Botucatu, Departamento de Genética, Rubião Júnior s/n, Botucatu, São Paulo, 18618-000, Brazil
2 - Embrapa - IAC, Centro de Café Alcides Carvalho, Av. Theodureto de Almeida Camargo, 1500, Campinas, São Paulo, 13001-970, Brazil
Presentation Type: Oral Paper:Papers for Sections
Date: Tuesday, August 1st, 2006
Time: 8:15 AM