title: Cloning and expression in Escherichia coli of the gene encoding an extracellular deoxyribonuclease (DNase) from Aeromonas hydrophila.
abstract: The gene encoding an extracellular DNase from Aeromonas hydrophila CHC-1 has been cloned and sequenced. Following expression of the dns in Escherichia coli, it was revealed that some of the cloned enzyme was present in the cell-free extracellular supernatant fluid, and there was no cell lysis and concurrent release of cytoplasmic or periplasmic proteins. Therefore, results suggest that E. coli cells were capable of secreting the DNase extracellularly, albeit very inefficiently. The dns is transcribed from its own promoter in E. coli, and expressed as a 25-kDa product, as determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis of the culture supernatant preparations followed by a DNA-hydrolysis assay. Nucleotide sequence analysis predicted a single open reading frame of 690 bp encoding a 230-amino acid (aa) polypeptide, with a potential 20-aa signal peptide located at the N terminus of the predicted protein. The deduced aa sequence of the entire protein is highly homologous with that of the DNase of Vibrio cholerae.
authors.affiliation: Department of Biochemistry, Medical College, National Cheng Kung University, Tainan, Taiwan.
authors.forename: M C
authors.initials: MC
authors.lastname: Chang
authors.forename: S Y
authors.initials: SY
authors.lastname: Chang
authors.forename: S L
authors.initials: SL
authors.lastname: Chen
authors.forename: S M
authors.initials: SM
authors.lastname: Chuang
mesh_terms: D016980:Aeromonas hydrophila / Q000201:enzymology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D003851:Deoxyribonucleases / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
chemical_list: D004269:DNA, Bacterial; D003851:Deoxyribonucleases