title: Evidence showing duplication and recombination of cel genes in tandem from hyperthermophilic Thermotoga sp.
abstract: This study was conducted to assess the gene duplication and diversification of tandem cellulase genes in thermophilic bacteria. The tandem cellulase genes cel5C and cel5D were cloned from Thermotoga maritima MSB8, and a survey of the thermophilic bacterial genome for tandem cel genes from the databases was carried out. A clone having 2.3 kb fragment from T. maritima MSB8 showed cellulase activity, which had two open reading frames in tandem (cel5C and cel5D). The cel5C gene has 954 bp, which encodes a protein of 317 amino acid residues with a signal peptide of 23 amino acids, and the other gene cel5D consisting of 990 bp encoding a protein of 329 amino acid residues. These two proteins have similarity with the enzymes of glycosyl hydrolase family 5. From the enzyme assay, it was observed that Cel5C was extracellular and Cel5D was intracellular cellulase. Phylogenetic and homology matrix analyses of DNA and protein sequences revealed that family 12 cellulase enzymes Cel12A and Cel12B displayed higher homology (>50 %), but Cel5C and Cel5D enzymes belong to family 5 displayed lower homology (<30 %). In addition, repeated and mirror sequences in tandem genes are supposed to show the existence of gene duplication and recombination.
authors.affiliation: Gyeongsangnam-do Agricultural Research and Extension Service, Chinju 660-360, Republic of Korea. goguma99@korea.kr
authors.forename: Min Keun
authors.initials: MK
authors.lastname: Kim
authors.forename: Tae Ho
authors.initials: TH
authors.lastname: Kang
authors.forename: Jungho
authors.initials: J
authors.lastname: Kim
authors.forename: Hoon
authors.initials: H
authors.lastname: Kim
authors.forename: Han Dae
authors.initials: HD
authors.lastname: Yun
mesh_terms: D044602:Cellulases / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D019143:Evolution, Molecular; D005110:Extracellular Space / Q000201:enzymology; D020440:Gene Duplication; D006026:Glycoside Hydrolases / Q000235:genetics; D042541:Intracellular Space / Q000201:enzymology; D008958:Models, Molecular; D010802:Phylogeny; D011487:Protein Conformation; D011995:Recombination, Genetic; D020124:Thermotoga maritima / Q000166:cytology / Q000201:enzymology* / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D044602:Cellulases; D006026:Glycoside Hydrolases