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Table 1 Strains, plasmids and primers used in this study

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

 
  1. 1Restriction sites are italicized.