The cloning and nucleotide sequence of the serine protease gene (aspA) of Aeromonas salmonicida ssp. salmonicida.

The gene for Aeromonas salmonicida serine protease has been cloned into phagemid pTZ18R in two restriction fragments, 2.0-kb PstI and 2.3-kb KpnI, of genomic DNA. The nucleotide sequences of the two fragments have been determined, in both directions, after subcloning, by double-stranded sequencing of nested deletions. An open reading frame of 1863 bp translated into a sequence of 621 amino acids, a 24-amino acid signal peptide and a 597-amino acid mature enzyme of molecular mass 64,173 Da. The consensus sequence, NGTS, of a serine protease substrate primary binding site was identified and a putative ribosome-binding site GGAG occurred 6 bp upstream of the ATG initiation codon.

[1]  B. M. Phipps,et al.  Structure of the tetragonal surface virulence array protein and gene of Aeromonas salmonicida. , 1991, The Journal of biological chemistry.

[2]  A. Ellis An appraisal of the extracellular toxins of Aeromonas salmonicida ssp. salmonicida , 1991 .

[3]  A. Driessen,et al.  The enzymology of protein translocation across the Escherichia coli plasma membrane. , 1991, Annual review of biochemistry.

[4]  J. Burke,et al.  Rapid isolation of miniprep DNA for double strand sequencing. , 1990, Nucleic acids research.

[5]  Sydney Brenner,et al.  The molecular evolution of genes and proteins: a tale of two serines , 1988, Nature.

[6]  G. Coleman,et al.  The combined effect of isolated Aeromonas salmonicida protease and haemolysin on Atlantic salmon, Salmo salar L., compared with that of a total extracellular products preparation , 1988 .

[7]  G. Coleman,et al.  Identification of major common extracellular proteins secreted by Aeromonas salmonicida strains isolated from diseased fish , 1987, Applied and environmental microbiology.

[8]  D. Mead,et al.  Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. , 1986, Protein engineering.

[9]  D. Glover DNA cloning : a practical approach , 1985 .

[10]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[11]  R. Doolittle,et al.  A simple method for displaying the hydropathic character of a protein. , 1982, Journal of molecular biology.

[12]  R. Roberts Microbial diseases of fish , 1982 .

[13]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[14]  E. Southern Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[15]  U. Schibler,et al.  Changes in size and secondary structure of the ribosomal transcription unit during vertebrate evolution. , 1975, Journal of molecular biology.

[16]  B. Witkop,et al.  Nonenzymatic cleavage of peptide bonds: the methionine residues in bovine pancreatic ribonuclease. , 1962, The Journal of biological chemistry.