A low-fibronectin-binding mutant of Staphylococcus aureus 879R4S has Tn918 inserted into its single fnb gene.
A low-fibronectin-binding mutant of Staphylococcus aureus strain 879R4SSp generated by transposon Tn918 mutagenesis is attenuated in a rat endocarditis model (J.M. Kuypers & R.A. Procter, 1989, Infect Immun 57, 2306-2312). PCR and Southern hybridization analysis with primers and probes, respectively, for the fnbA and fnbB genes of strains 8325-4 showed that strain 879R4SSp possesses a single fnb gene which is homologous to fnbA. This was confirmed by sequencing 41 bp of 5' non-coding and 237 bp of 5' coding DNA, which showed 97% base identity to fnbA. Southern hybridization and sequencing showed that Tn918 was inserted 41 bp 5' to fnbA in the mutant 879R4SSp/1536, between the promoter and initiation codon. Reduced adherence of the mutant to surface-bound fibronectin correlated with lower expression of a 180 kDa wall-associated fibronectin-binding protein.
Greene
C
C
Microbiology Department, Moyne Institute of Preventive Medicine, Trinity College, Dublin, Ireland.
Vaudaux
P E
PE
Francois
P
P
Proctor
R A
RA
McDevitt
D
D
Foster
T J
TJ
D018829:Adhesins, Bacterial; D000818:Animals; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D001426:Bacterial Proteins; D001483:Base Sequence; D015139:Blotting, Southern; D002352:Carrier Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; D004696:Endocarditis / Q000382:microbiology; D004926:Escherichia coli; D005353:Fibronectins / Q000378:metabolism*; D005838:Genotype; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D051381:Rats; D015183:Restriction Mapping; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000235:genetics / Q000502:physiology*
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; D005353:Fibronectins; D015345:Oligonucleotide Probes; C078635:fibronectin-binding proteins, bacterial