GeneCARD-FISH: detection of tceA and vcrA reductive dehalogenase genes in Dehalococcoides mccartyi by fluorescence in situ hybridization.
作者： B. Matturro , S. Rossetti
DOI: 10.1016/J.MIMET.2015.01.005
关键词:
摘要: Due to the direct involvement in biodegradation of chlorinated solvents, reductive dehalogenase genes (RDase) are considered biomarkers metabolic potential different strains Dehalococcoides mccartyi (Dhc). This is known be only microbe able completely reduce toxic solvents harmless ethene. In last years, several Molecular Biological Tools (MBTs) have been developed optimize detectability Dhc cells and/or RDase genes, with particular attention most important indicators ethene formation, namely tceA and vcrA genes. Despite qPCR has indicated as MBT choice, use CARD-FISH recently demonstrated provide a more accurate quantification wide concentration range, overcoming drawbacks loosing nucleic acids during preparation sample associated qPCR. assays usually target 16S rRNA up date no protocol discriminate by detecting developed. study reports first evidence situ detection into applying new procedure named geneCARD-FISH. carrying were identified quantified PCE-to-ethene dechlorinating microbial enrichment overall they represented 58.63%±2.45% 40.46%±1.86% total cells, respectively. These values markedly higher than those obtained qPCR, which strongly underestimated actual gene (0.08%±0.01% copies). The assay was successfully applied also for analysis environmental samples remarkably strengthens biomonitoring activities at field scale providing specific discrimination key-RDase
elsevier.com
sci-hub.se 参考文章(34)
Cristina Moraru, Phyllis Lam, Bernhard M. Fuchs, Marcel M. M. Kuypers, Rudolf Amann, GeneFISH--an in situ technique for linking gene presence and cell identity in environmental microorganisms. Environmental Microbiology. ,vol. 12, pp. 3057- 3073 ,(2010) , 10.1111/J.1462-2920.2010.02281.X
Cristina Moraru, Gabriel Moraru, Bernhard M. Fuchs, Rudolf Amann, Concepts and software for a rational design of polynucleotide probes Environmental Microbiology Reports. ,vol. 3, pp. 69- 78 ,(2011) , 10.1111/J.1758-2229.2010.00189.X
Lorenz Adrian, Ulrich Szewzyk, Jörg Wecke, Helmut Görisch, Bacterial dehalorespiration with chlorinated benzenes Nature. ,vol. 408, pp. 580- 583 ,(2000) , 10.1038/35046063
Chang Ding, Jianzhong He, Molecular techniques in the biotechnological fight against halogenated compounds in anoxic environments Microbial Biotechnology. ,vol. 5, pp. 347- 367 ,(2012) , 10.1111/J.1751-7915.2011.00313.X
Tina Hölscher, Rosa Krajmalnik-Brown, Kirsti M. Ritalahti, Friedrich von Wintzingerode, Helmut Görisch, Frank E. Löffler, Lorenz Adrian, Multiple Nonidentical Reductive-Dehalogenase-Homologous Genes Are Common in Dehalococcoides Applied and Environmental Microbiology. ,vol. 70, pp. 5290- 5297 ,(2004) , 10.1128/AEM.70.9.5290-5297.2004
Ruth Ellen Richardson, Genomic insights into organohalide respiration. Current Opinion in Biotechnology. ,vol. 24, pp. 498- 505 ,(2013) , 10.1016/J.COPBIO.2013.02.014
A JUSTE, B THOMMA, B LIEVENS, Recent advances in molecular techniques to study microbial communities in food-associated matrices and processes Food Microbiology. ,vol. 25, pp. 745- 761 ,(2008) , 10.1016/J.FM.2008.04.009
Mauro Majone, Roberta Verdini, Federico Aulenta, Simona Rossetti, Valter Tandoi, Nicolas Kalogerakis, Spiros Agathos, Sebastià Puig, Giulio Zanaroli, Fabio Fava, In situ groundwater and sediment bioremediation: barriers and perspectives at European contaminated sites New Biotechnology. ,vol. 32, pp. 133- 146 ,(2015) , 10.1016/J.NBT.2014.02.011
Shuji Kawakami, Kengo Kubota, Hiroyuki Imachi, Takashi Yamaguchi, Hideki Harada, Akiyoshi Ohashi, Detection of single copy genes by two-pass tyramide signal amplification fluorescence in situ hybridization (Two-Pass TSA-FISH) with single oligonucleotide probes. Microbes and Environments. ,vol. 25, pp. 15- 21 ,(2010) , 10.1264/JSME2.ME09180
Hans-Joachim Höltke, Christoph Kessler, Non-radioactive labeling of RNA transcripts in vitro with the hapten digoxigenin (DIG); hybridization and ELISA-based detection Nucleic Acids Research. ,vol. 18, pp. 5843- 5851 ,(1990) , 10.1093/NAR/18.19.5843