High-throughput PCR screening of genes for three-component regulatory system putatively involved in quorum sensing from low-G+C gram-positive bacteria

Jiro Nakayama, Antoon D.L. Akkermans, Willem M. De Vos

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Quorum sensing of Gram-positive bacteria is often regulated by three-component regulatory system composed of autoinducing peptide, sensor kinase and response regulator. We used PCR to study a gene cassette encoding this three-component regulatory system. Degenerate primers were designed from consensus amino acid sequences in the HPK10 subfamily, mostly involved in quorum sensing. Products amplified from genomic DNA of Lactobacillus, Enterococcus, and Clostridium species were cloned and sequenced; their deduced amino acid sequences were similar to those of members of the HPK10 subfamily. Complete genes for the putative gene cassette were cloned by inverse PCR from L. paracasei E93490 and L. plantarum WCFS6. Phylogenetic analysis grouped the cloned putative HPKs into the HPK10 subfamily. These results indicated the usefulness of this high-throughput gene screening and suggested that the three-component regulatory gene cassette are widely present.

Original languageEnglish
Pages (from-to)480-489
Number of pages10
JournalBioscience, Biotechnology and Biochemistry
Volume67
Issue number3
DOIs
Publication statusPublished - Jan 1 2003

Fingerprint

Gene Components
Quorum Sensing
Gram-Positive Bacteria
Bacteria
Screening
Genes
Throughput
Polymerase Chain Reaction
Amino Acid Sequence
Clostridium
Enterococcus
Lactobacillus
Regulator Genes
Amino Acids
Phosphotransferases
Peptides
DNA
Sensors

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

High-throughput PCR screening of genes for three-component regulatory system putatively involved in quorum sensing from low-G+C gram-positive bacteria. / Nakayama, Jiro; Akkermans, Antoon D.L.; De Vos, Willem M.

In: Bioscience, Biotechnology and Biochemistry, Vol. 67, No. 3, 01.01.2003, p. 480-489.

Research output: Contribution to journalArticle

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