Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus.
We have previously identified a gene in Staphylococcus aureus, agr, whose activity is required for high-level post-exponential-phase expression of a series of secreted proteins. In this paper, we describe the cloning of this gene in Escherichia coli by using an inserted transposon (Tn551) as a cloning probe. The cloned gene, consisting of a 241-codon open reading frame containing the site of the transposon insertion, was recloned to an S. aureus vector, pSK265, and shown to be functional in S. aureus. Activity was evaluated by determinations of alpha-hemolysin, beta-hemolysin, and toxic shock syndrome toxin-1 production in early-stationary-phase cultures. The cloned gene showed considerable variation with respect to different exoproteins and different host strains compared with the chromosomal agr determinant; this variation could not be attributed to the higher copy number of the cloned gene and probably reflects inapparent subtleties of the regulatory system.
Peng
H L
HL
Public Health Research Institute, New York, New York 10016.
Novick
R P
RP
Kreiswirth
B
B
Kornblum
J
J
Schlievert
P
P
D000595:Amino Acid Sequence; D001427:Bacterial Toxins; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004768:Enterotoxins / Q000096:biosynthesis; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005822:Genetic Vectors; D006460:Hemolysin Proteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009154:Mutation; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000235:genetics; D013211:Staphylococcus aureus / Q000235:genetics*; D018089:Superantigens; D014158:Transcription, Genetic
D001427:Bacterial Toxins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004768:Enterotoxins; D006460:Hemolysin Proteins; D012329:RNA, Bacterial; D018089:Superantigens; C030107:enterotoxin F, Staphylococcal; D004262:DNA Restriction Enzymes