A new set of reference genes for RT-qPCR assays in the yeast Dekkera bruxellensis
| Autor: | Gonçalo Amarante Guimarães Pereira, Marcos Antonio de Morais, Will de Barros Pita, Luciana Filgueira Pereira, Fernanda Cristina Bezerra Leite, Anna Theresa de Souza Liberal, Marcelo Falsarella Carazzolle |
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| Rok vydání: | 2012 |
| Předmět: |
Genetics
Regulation of gene expression Normalization (statistics) Dekkera Genes Fungal Immunology Reproducibility of Results General Medicine Cellobiose Biology Real-Time Polymerase Chain Reaction Applied Microbiology and Biotechnology Microbiology Yeast chemistry.chemical_compound Real-time polymerase chain reaction chemistry Gene Expression Regulation Fungal Reference genes Gene expression RNA Messenger Molecular Biology Gene DNA Primers |
| Zdroj: | Canadian Journal of Microbiology. 58:1362-1367 |
| ISSN: | 1480-3275 0008-4166 |
| DOI: | 10.1139/cjm-2012-0457 |
| Popis: | The yeast Dekkera bruxellensis has been recently regarded as an important microorganism for bioethanol production owing to its ability to convert glucose, sucrose, and cellobiose to ethanol. The aim of this work was to validate a new set of reference genes for gene expression analysis by quantitative real-time PCR in D. bruxellensis and compare the influence of the method of choice for quantification of mRNA levels with the reliability of our data. Three candidate reference genes, DbEFA1, DbEFB1, and DbYNA1, were used in a quantitative analysis of 4 genes of interest, DbYNR1, DbTPS1, DbADH7, and DbUBA4, based on an approach for calculating the normalization factors by means of the geNorm applet. Each reference gene was also individually used for a [Formula: see text] (comparative Cq method) calculation of the relative expression of genes of interest. Our results showed that the 3 reference genes provided enough stability and were complementary to the normalization factors method in different culture conditions. This work was able to confirm the usefulness of a previously reported reference gene, EFA1/TEF1, and increased the set of possible reference genes in D. bruxellensis to 4. Moreover, this can improve the reliability of the analysis of the regulation of gene expression in the industrial yeast D. bruxellensis. |
| Databáze: | OpenAIRE |
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