A new set of reference genes for RT-qPCR assays in the yeast Dekkera bruxellensis

被引：14
作者：
Pita, Will de Barros [1 ]
Bezerra Leite, Fernanda Cristina [1 ]
de Souza Liberal, Anna Theresa [1 ]
Pereira, Luciana Filgueira [1 ]
Carazzolle, Marcelo Falsarella [2 ]
Pereira, Goncalo Amarante [2 ]
de Morais, Marcos Antonio, Jr. [1 ,3 ]
机构：
[1] Univ Fed Pernambuco, Interdept Res Grp Metab Engn, BR-50670901 Recife, PE, Brazil
[2] Univ Estadual Campinas, Lab Genom & Express, BR-13083970 Campinas, SP, Brazil
[3] Univ Fed Pernambuco, Dept Genet, BR-50670901 Recife, PE, Brazil
来源：
CANADIAN JOURNAL OF MICROBIOLOGY | 2012年 / 58卷 / 12期
关键词：
Dekkera bruxellensis; gene expression; geNorm; reference genes; RT-qPCR; SACCHAROMYCES-CEREVISIAE; PCR; ABILITY;
D O I：
10.1139/cjm-2012-0457
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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 2(-Delta Delta Cq) (comparative C-q 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.
引用
收藏
页码：1362 / 1367
页数：6
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共 13 条
  • [1] Growth rates of Dekkera/Brettanomyces yeasts hinder their ability to compete with Saccharomyces cerevisiae in batch corn mash fermentations
    Abbott, DA
    Hynes, SH
    Ingledew, WM
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2005, 66 (06) : 641 - 647
  • [2] Detection and identification of wild yeast contaminants of the industrial fuel ethanol fermentation process
    Basillo, A. C. M.
    de Araujo, P. R. L.
    de Morais, J. O. F.
    da Silva Filho, E. A.
    de Morais, M. A., Jr.
    Simoes, D. A.
    [J]. CURRENT MICROBIOLOGY, 2008, 56 (04) : 322 - 326
  • [3] Fermentation characteristics of Dekkera bruxellensis strains
    Blomqvist, Johanna
    Eberhard, Thomas
    Schnurer, Johan
    Passoth, Volkmar
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2010, 87 (04) : 1487 - 1497
  • [4] The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments
    Bustin, Stephen A.
    Benes, Vladimir
    Garson, Jeremy A.
    Hellemans, Jan
    Huggett, Jim
    Kubista, Mikael
    Mueller, Reinhold
    Nolan, Tania
    Pfaffl, Michael W.
    Shipley, Gregory L.
    Vandesompele, Jo
    Wittwer, Carl T.
    [J]. CLINICAL CHEMISTRY, 2009, 55 (04) : 611 - 622
  • [5] The ability to use nitrate confers advantage to Dekkera bruxellensis over S. cerevisiae and can explain its adaptation to industrial fermentation processes
    de Barros Pita, Will
    Bezerra Leite, Fernanda Cristina
    de Souza Liberal, Anna Theresa
    Simoes, Diogo Ardaillon
    de Morais, Marcos Antonio, Jr.
    [J]. ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY, 2011, 100 (01): : 99 - 107
  • [6] Identification of Dekkera bruxellensis as a major contaminant yeast in continuous fuel ethanol fermentation
    Liberal, A. T. de Souza
    Basilio, A. C. M.
    Resende, A. do Monte
    Brasileiro, B. T. V.
    da Silva-Filho, E. A.
    de Morais, J. O. F.
    Simoes, D. A.
    de Morais, M. A., Jr.
    [J]. JOURNAL OF APPLIED MICROBIOLOGY, 2007, 102 (02) : 538 - 547
  • [7] Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method
    Livak, KJ
    Schmittgen, TD
    [J]. METHODS, 2001, 25 (04) : 402 - 408
  • [8] Adaptation of yeasts Saccharomyces cerevisiae and Brettanomyces bruxellensis to winemaking conditions: a comparative study of stress genes expression
    Nardi, Tiziana
    Remize, Fabienne
    Alexandre, Herve
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2010, 88 (04) : 925 - 937
  • [9] Dekkera bruxellensis and Lactobacillus vini form a stable ethanol-producing consortium in a commercial alcohol production process
    Passoth, Volkmar
    Blomqvist, Johanna
    Schnurer, Johan
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (13) : 4354 - 4356
  • [10] Mousy off-flavor: A review
    Snowdon, Eleanor M.
    Bowyer, Michael C.
    Grbin, Paul R.
    Bowyer, Paul K.
    [J]. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2006, 54 (18) : 6465 - 6474
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