[Cloning and sequence analysis of the gene encoding NiFe-hydrogenase from Klebsiella pneumoniae].
Degenerate PCR primers were designed by multiple alignment of the protein sequences of known structural genes encoding the catalytic subunits of NiFe-hydrogenases obtained from Swiss-Prot Protein Sequence Database through CLUSTAL-W software and compared for conserved sequence motifs. An amplified PCR product 1 kb in size was obtained from the genomic DNA of Klebsiella pneumoniae using a set of degenerate primers, and then inverse PCR technique was used to obtain the full hydrogenase coding region. A predicted secondary structure and 3D structural model were constructed by homology modeling and docking. On the basis of these results, it was inferred that NiFe-hydrogenase from Klebsiella pneumoniae belongs to the membrane-bound H2 evolving hydrogenase group (Ech hydrogenase group).
Liu
Fei
F
Province Key Laboratory of Industrial Biotechnology( Huaqiao University), Quanzhou 362021, China.
Fang
Bai-Shan
BS
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D030562:Databases, Protein; D006860:Hydrogen Bonding; D006864:Hydrogenase / Q000737:chemistry / Q000235:genetics*; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; C043296:nickel-iron hydrogenase; D006864:Hydrogenase