Selection of housekeeping genes for normalization by real-time RT-PCR: analysis of Or-MYB1 gene expression in Orobanche ramosa development
| Autor: | Belen Roman, Jose V. Die, M. T. Moreno, Salvador Nadal, C. I. González-Verdejo, A. Jiménez-Marín |
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| Rok vydání: | 2008 |
| Předmět: |
Genetics
Candidate gene Time Factors biology Orobanche Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling Biophysics Cell Biology Reference Standards biology.organism_classification Genes Plant Biochemistry Sensitivity and Specificity Housekeeping gene Gene expression profiling Gene Expression Regulation Plant Reference genes Gene expression RNA Messenger Molecular Biology Transcription factor Gene Orobanche ramosa |
| Zdroj: | Analytical biochemistry. 379(2) |
| ISSN: | 1096-0309 |
| Popis: | Real-time PCR has become the method of choice for accurate and in-depth expression studies of candidate genes. To avoid bias, real-time PCR is referred to one or several internal control genes that should not fluctuate among treatments. A need for reference genes in the parasitic plant Orobanche ramosa has emerged, and the studies in this area have not yet been evaluated. In this study, the genes 18S rRNA, Or-act1, Or-tub1, and Or-ubq1 were compared in terms of expression stability using the BestKeeper software program. Among the four common endogenous control genes, Or-act1 and Or-ubq1 were the most stable in O. ramosa samples. In parallel, a study was carried out studying the expression of the transcription factor Or-MYB1 that seemed to be implicated during preinfection stages. The normalization strategy presented here is a prerequisite to accurate real-time PCR expression profiling that, among other things, opens up the possibility of studying messenger RNA levels of low-copy-number-like transcription factors. |
| Databáze: | OpenAIRE |
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