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Nucleic Acids Res. 1986 Jul 25; 14(14): 5843–5855.
PMCID: PMC311595

Cloning and sequencing of Serratia protease gene.

Abstract

The gene encoding an extracellular metalloproteinase from Serratia sp. E-15 has been cloned, and its complete nucleotide sequence determined. The amino acid sequence deduced from the nucleotide sequence reveals that the mature protein of the Serratia protease consists of 470 amino acids with a molecular weight of 50,632. The G+C content of the coding region for the mature protein is 58%; this high G+C content is due to a marked preference for G+C bases at the third position of the codons. The gene codes for a short pro-peptide preceding the mature protein. The Serratia protease gene was expressed in Escherichia coli and Serratia marcescens; the former produced the Serratia protease in the cells and the latter in the culture medium. Three zinc ligands and an active site of the Serratia protease were predicted by comparing the structure of the enzyme with those of thermolysin and Bacillus subtilis neutral protease.

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Selected References

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  • Katsuya Y, Hamada K, Hata Y, Tanaka N, Sato M, Katsube Y, Kakiuchi K, Miyata K. Preliminary X-ray studies on Serratia protease. J Biochem. 1985 Oct;98(4):1139–1142. [PubMed]
  • Lee IS, Wakabayashi S, Miyata K, Tomoda K, Yoneda M, Kangawa K, Minamino N, Matsuo H, Matsubara H. Serratia protease. Amino acid sequences of both termini, the 53 residues in the middle region containing the sole methionine residue, and a probable zinc-binding region. J Biochem. 1984 Nov;96(5):1409–1418. [PubMed]
  • Schmitz G, Braun V. Cell-bound and secreted proteases of Serratia marcescens. J Bacteriol. 1985 Mar;161(3):1002–1009. [PMC free article] [PubMed]
  • Kaplan JB, Merkel WK, Nichols BP. Evolution of glutamine amidotransferase genes. Nucleotide sequences of the pabA genes from Salmonella typhimurium, Klebsiella aerogenes and Serratia marcescens. J Mol Biol. 1985 Jun 5;183(3):327–340. [PubMed]
  • Jones JD, Grady KL, Suslow TV, Bedbrook JR. Isolation and characterization of genes encoding two chitinase enzymes from Serratia marcescens. EMBO J. 1986 Mar;5(3):467–473. [PMC free article] [PubMed]
  • Vasantha N, Thompson LD, Rhodes C, Banner C, Nagle J, Filpula D. Genes for alkaline protease and neutral protease from Bacillus amyloliquefaciens contain a large open reading frame between the regions coding for signal sequence and mature protein. J Bacteriol. 1984 Sep;159(3):811–819. [PMC free article] [PubMed]
  • Yang MY, Ferrari E, Henner DJ. Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived deletion mutation. J Bacteriol. 1984 Oct;160(1):15–21. [PMC free article] [PubMed]
  • Messing J, Crea R, Seeburg PH. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. [PMC free article] [PubMed]
  • LENNOX ES. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. [PubMed]
  • Lovett PS, Keggins KM. Bacillus subtilis as a host for molecular cloning. Methods Enzymol. 1979;68:342–357. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Burnette WN. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. [PubMed]
  • POLLOCK MR, RICHMOND MH. Low cyst(e)ine content of bacterial extracellular proteins: its possible physiological significance. Nature. 1962 May 5;194:446–449. [PubMed]
  • Weaver LH, Kester WR, Matthews BW. A crystallographic study of the complex of phosphoramidon with thermolysin. A model for the presumed catalytic transition state and for the binding of extended substances. J Mol Biol. 1977 Jul;114(1):119–132. [PubMed]
  • Levy PL, Pangburn MK, Burstein Y, Ericsson LH, Neurath H, Walsh KA. Evidence of homologous relationship between thermolysin and neutral protease A of Bacillus subtilis. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4341–4345. [PMC free article] [PubMed]
  • Chou PY, Fasman GD. Prediction of protein conformation. Biochemistry. 1974 Jan 15;13(2):222–245. [PubMed]
  • Wells JA, Ferrari E, Henner DJ, Estell DA, Chen EY. Cloning, sequencing, and secretion of Bacillus amyloliquefaciens subtilisin in Bacillus subtilis. Nucleic Acids Res. 1983 Nov 25;11(22):7911–7925. [PMC free article] [PubMed]
  • Jacobs M, Eliasson M, Uhlén M, Flock JI. Cloning, sequencing and expression of subtilisin Carlsberg from Bacillus licheniformis. Nucleic Acids Res. 1985 Dec 20;13(24):8913–8926. [PMC free article] [PubMed]
  • Felmlee T, Pellett S, Lee EY, Welch RA. Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J Bacteriol. 1985 Jul;163(1):88–93. [PMC free article] [PubMed]
  • Felmlee T, Pellett S, Welch RA. Nucleotide sequence of an Escherichia coli chromosomal hemolysin. J Bacteriol. 1985 Jul;163(1):94–105. [PMC free article] [PubMed]

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