The use of degenerate, sensor gene-specific, oligodeoxyribonucleotide primers to amplify DNA fragments from Staphylococcus aureus

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Abstract

The sensor proteins of bacterial two-component regulatory systems comprise a large family of proteins that are involved in environmental sensing and signal transduction. To study these proteins in the Gram+ pathogen, Staphylococcus aureus, two pairs of degenerate oligodeoxyribonucleotides (oligos) that corresponded to conserved sequences contained within sensor protein-encoding genes were synthesized. Using these oligo primers, DNA fragments from S. aureus were amplified by polymerase chain reaction (PCR), cloned in Escherichia coli, and sequenced. Comparison of the deduced amino acid sequences from these cloned fragments to the sequences contained in the GenBank database suggest that some of the PCR products were derived from sensor protein-encoding genes. However, several other fragments were identified that encoded peptides with up to 65% identity to transport proteins. Given the biochemical and functional properties of some of these proteins, these data suggest that sensor and transport proteins may be evolutionarily related.

Original languageEnglish (US)
Pages (from-to)99-103
Number of pages5
JournalGene
Volume123
Issue number1
DOIs
StatePublished - Jan 15 1993

Fingerprint

DNA Primers
Staphylococcus aureus
DNA
Genes
Proteins
Carrier Proteins
Polymerase Chain Reaction
Bacterial Proteins
Conserved Sequence
Oligodeoxyribonucleotides
Nucleic Acid Databases
Amino Acid Sequence
Signal Transduction
Databases
Escherichia coli
Peptides

Keywords

  • PCR
  • Two-component regulatory systems
  • environmental stimuli
  • sensor protein
  • signal transduction
  • transport

ASJC Scopus subject areas

  • Genetics

Cite this

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abstract = "The sensor proteins of bacterial two-component regulatory systems comprise a large family of proteins that are involved in environmental sensing and signal transduction. To study these proteins in the Gram+ pathogen, Staphylococcus aureus, two pairs of degenerate oligodeoxyribonucleotides (oligos) that corresponded to conserved sequences contained within sensor protein-encoding genes were synthesized. Using these oligo primers, DNA fragments from S. aureus were amplified by polymerase chain reaction (PCR), cloned in Escherichia coli, and sequenced. Comparison of the deduced amino acid sequences from these cloned fragments to the sequences contained in the GenBank database suggest that some of the PCR products were derived from sensor protein-encoding genes. However, several other fragments were identified that encoded peptides with up to 65{\%} identity to transport proteins. Given the biochemical and functional properties of some of these proteins, these data suggest that sensor and transport proteins may be evolutionarily related.",
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AB - The sensor proteins of bacterial two-component regulatory systems comprise a large family of proteins that are involved in environmental sensing and signal transduction. To study these proteins in the Gram+ pathogen, Staphylococcus aureus, two pairs of degenerate oligodeoxyribonucleotides (oligos) that corresponded to conserved sequences contained within sensor protein-encoding genes were synthesized. Using these oligo primers, DNA fragments from S. aureus were amplified by polymerase chain reaction (PCR), cloned in Escherichia coli, and sequenced. Comparison of the deduced amino acid sequences from these cloned fragments to the sequences contained in the GenBank database suggest that some of the PCR products were derived from sensor protein-encoding genes. However, several other fragments were identified that encoded peptides with up to 65% identity to transport proteins. Given the biochemical and functional properties of some of these proteins, these data suggest that sensor and transport proteins may be evolutionarily related.

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