title: Genetic diversity and amplification of different clostridial [FeFe] hydrogenases by group-specific degenerate primers.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Centre for Protein Engineering, Bacterial Physiology and Genetics, University of Liège, Liège, Belgium. mcalusinska@ulg.ac.be
authors.forename: M
authors.initials: M
authors.lastname: Calusinska
authors.forename: B
authors.initials: B
authors.lastname: Joris
authors.forename: A
authors.initials: A
authors.lastname: Wilmotte
mesh_terms: D003013:Clostridium / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry*; D014644:Genetic Variation; D006801:Humans; D006864:Hydrogenase / Q000145:classification / Q000235:genetics*; D007506:Iron-Sulfur Proteins / Q000145:classification / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D007506:Iron-Sulfur Proteins; C118830:iron hydrogenase; D006864:Hydrogenase