ORBi: Calusinska Magdalena - Genetic diversity and amplification of different clostridial [FeFe] hydrogenases by group-specific degenerate primers

Reference : Genetic diversity and amplification of different clostridial [FeFe] hydrogenases by grou...


	Document type :	Scientific journals : Article
	Discipline(s) :	Life sciences : Biochemistry, biophysics & molecular biology
	To cite this reference:	http://hdl.handle.net/2268/101255

Title :	Genetic diversity and amplification of different clostridial [FeFe] hydrogenases by group-specific degenerate primers
Language :	English
Alternative title :	[fr] Diversité génétique et amplification de différentes hydrogénases (FeFe) de clostridies grâce à des amorces dégénérées spécifiques de groupes
Author, co-author :	Calusinska, Magdalena [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
	Joris, Bernard [Université de Liège - ULg > Département des sciences de la vie > Physiologie et génétique bactériennes >]
	Wilmotte, Annick [Université de Liège - ULg > Département des sciences de la vie > Physiologie et génétique bactériennes >]
Publication date :	6-Aug-2011
Journal title :	Letters in Applied Microbiology
Publisher :	Blackwell Publishing
Volume :	53
Pages :	473-480
Peer reviewed :	Yes (verified by ORBi)
Audience :	International
ISSN :	0266-8254
e-ISSN :	1472-765X
Keywords :	[en] degenerate primers ; [FeFe] hydrogenase ; clostridium
Abstract :	[en] Aims: The aim of this study was to explore and characterize the genetic diversity of [FeFe] hydrogenases in a representative set of strains from Clostridium sp. and to reveal the existence of neither yet detected nor characterized [FeFe] hydrogenases in hydrogen-producing strains. Methods and Results: The genomes of 57 Clostridium strains (34 different genotypic species), representing six phylogenetic clusters based on their 16S rRNA sequence analysis (cluster I, III, XIa, XIb, XIV and XVIII), were screened for different [FeFe] hydrogenases. Based on the obtained alignments, ten pairs of [FeFe] hydrogenase cluster-specific degenerate primers were newly designed. Ten Clostridium strains were screened by PCRs to assess the specificity of the primers designed and to examine the genetic diversity of [FeFe] hydrogenases. Using this approach, a diversity of hydrogenase genes was discovered in several species previously shown to produce hydrogen in bioreactors: Clostridium sartagoforme, Clostridium felsineum, Clostridium roseum and Clostridium pasteurianum. Conclusions: The newly designed [FeFe] hydrogenase cluster-specific primers, targeting the cluster-conserved regions, allow for a direct amplification of a specific hydrogenase gene from the species of interest. Significance and Impact of the Study: Using this strategy for a screening of different Clostridium ssp. will provide new insights into the diversity of hydrogenase genes and should be a first step to study a complex hydrogen metabolism of this genus.
Research units :	Centre d'Ingénierie des Protéines - CIP
Funders :	Communauté française de Belgique - CfB
Name of the research project :	Micro-H2
Target :	Researchers ; Professionals ; Students
Permalink :	http://hdl.handle.net/2268/101255
Mentions required by the publisher for OA :	Pas d'OA possible

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