From: Investigating the genetic regulation of the ECF sigma factor σS in Staphylococcus aureus
Strain, plasmid or primer | Genotype or description | Reference or source |
---|---|---|
E. coli | ||
DH5α | φ80 Δ(lacZ)M15 Δ(argF-lac)U169 endA1 recA1 | [12] |
hsdR17 (rK -mK +) deoR thi-1 supE44 gyrA96 relA1 | ||
S. aureus | ||
RN4220 | Restriction deficient transformation recipient | Lab stocks |
8325-4 | Wild-Type Laboratory Strain rsbU mutant | Lab stocks |
SH1000 | Wild-Type Laboratory Strain rsbU functional | [13] |
LES57 | SH1000 pAZ106::sigS-lacZ sigS + | 7 |
HKM08 | 8325-4 pAZ106::sigS-lacZ sigS + | This study |
HKM15 | LES57 isolate subject to random mutagenesis using nitrosoguanidine | This study |
HKM16 | LES57 isolate subject to random mutagenesis using nitrosoguanidine | This study |
HKM17 | 8325-4 pSC-A::tet::sigS-lacZ sigS + | This study |
HKM18 | 8325-4 pSC-A::tet::sigS-lacZ sigS + pRN3208 | This study |
Plasmids | ||
pSC-A | TA clone vector lacking Gram-positive origin of replication | Strata clone |
pRN3208 | TS shuttle vector harboring Tn551 | 15 |
pDG1515 | Tetracycline resistance antibiotic cassette in Bluescript KS(+) (Ampr) | [14] |
pLES205 | pAZ106 containing a 1. 4 kb sigS fragment | 7 |
pHKM4 | pSC-A containing the sigS-lacZ fragment from pLES205 | This study |
pHKM5 | pSC-A containing a tet cassette and sigS-lacZ fragment from pLES205 | This study |
Primers1 | ||
OL281 | ACT GGA TCC CAG TTG CAG ATG CAT CTC TCC | |
OL388 | GTT GTC TGA ATA AAT CGA TAA GG | |
OL522 | ATG TCT AGA GAG TAA TGC TAA CAT AGC | |
OL523 | ATG TCT AGA CCC AAA GTT GAT CCC TTA ACG | |
OL909 | ATG CTG CAG CAG GAC CCA ACG CTG CCC GAG | |
OL1039 | [Biotin]CGT GCC TTC AAT TTG ACC ATC ACG | |
OL1184 | AGC CGA CCT GAG AGG GTG A | |
OL1185 | TCT GGA CCG TGT CTC AGT TCC | |
OL1471 | TTT ATG GTA CCA TTT CAT TTT CCT GCT TTT TC | |
OL1562 | [biotin]GTA ATC CAT TGT TAC CTC CCG | |
OL1563 | [biotin] GTG GTG TTT GTT GTA TAC GTC | |
OL1568 | CGA TTA CGC AAA TGA ATG | |
OL1569 | CAA GTA GTC ATT CTC CAA G | |
OL1942 | GTA ATC CAT TGT TAC CTC CCG | |
OL1943 | GTG GTG TTT GTT GTA TAC GTC | |
OL2967 | [biotin]GGC TTT CAA TTT GAT TAC GTT T | |
OL2968 | [biotin]CTA AAT TAA AAG TAT AAC TGC ATT G | |
OL2979 | [biotin]GTA CAC CTC ATA TTA CGA CTT TTT C | |
OL2980 | [biotin]CAT TAG TGA GAA TCA TTG TCA ATT AG | |
OL3012 | [biotin]CCA TGA TAA CCC TCA CTT AAT ATA | |
OL3013 | [biotin]GAC ATA ACC TTC ACC TCG ATA GCA | |
OL3014 | CCA TGA TAA CCC TCA CTT AAT ATA | |
OL3015 | GAC ATA ACC TTC ACC TCG ATA GCA | |
OL3016 | [biotin]GCA TCT GAC ACA CAA GTA TTT GTG TTG | |
OL3017 | [biotin]CCG AGT CTG TCT TTA ACA CTG | |
OL3018 | GCA TCT GAC ACA CAA GTA TTT GTG TTG | |
OL3019 | CCG AGT CTG TCT TTA ACA CTG |