Cloning and sequence analysis of the gene encoding NiFe-hydrogenase from Klebsiella pneumoniae.
- Author:
Fei LIU
1
;
Bai-Shan FANG
Author Information
1. Province Key Laboratory of Industrial Biotechnology( Huaqiao University), Quanzhou 362021, China.
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
chemistry;
genetics;
Cloning, Molecular;
Codon;
genetics;
DNA, Bacterial;
chemistry;
genetics;
Databases, Protein;
Hydrogen Bonding;
Hydrogenase;
chemistry;
genetics;
Klebsiella pneumoniae;
enzymology;
genetics;
Models, Molecular;
Molecular Sequence Data;
Protein Conformation;
Protein Structure, Secondary;
Protein Structure, Tertiary;
Sequence Analysis, DNA
- From:
Chinese Journal of Biotechnology
2007;23(1):133-137
- CountryChina
- Language:Chinese
-
Abstract:
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).
