Table 1. Bacterial strains and plasmids used in this study.
Bacterial strain or plasmid | Descriptiona | Reference or source |
Strains | ||
Escherichia coli | ||
Alpha-Select Gold | F- deoR endA1 recA1 relA1 gyrA96 hsdR17(rk -, mk +) supE44 thi-1 phoA Δ(lacZYA-argF)U169 Φ80lacZΔM15 λ- | Bioline |
BL21(DE3) | F– ompT hsdSB(rB–, mB–) gal dcm (DE3) | Bioline |
Porphyromonas gingivalis | ||
ATCC 33277 | Wild-type | Oral Health CRC |
ECR455 | P. gingivalis 33277 har::ermF, Emr | This study |
ECR475 | ECR455 ermF::har cepA, Apr | This study |
Plasmids | ||
pBluescript II | Cloning vector; Apr | Stratagene |
pGEM-TEasy | Cloning vector linearized with T overhangs for ligation of PCR products generated with A overhangs by Taq polymerase; Apr | Promega |
pGEX-4T-Har | pGEX-4T-1 containing har and an engineered Factor Xa cleavage site, for cleavage of the expressed full-length Har from an N-terminal GST tag. | This study |
pGEX-4T-Har150 | pGEX-4T-1 containing a 3′ shortened har gene and an engineered Factor Xa cleavage site, for cleavage of the expressed C-terminally truncated Har150 from an N-terminal GST tag. | This study |
pGEX-4T-HarC97A | pGEX-4T-Har which had the TGT codon for Cys97 mutated to GCA for Ala. | This study |
pVA2198 | E. coli-Bacteroides shuttle vector carrying ermF-ermAM cassette; Emr | [76] |
pHarSOE1-4 | Recombination cassette for the deletion of har from P. gingivalis 33277, cloned blunt into the SmaI site of pBluescript II; Apr in E. coli | This study |
pEC474 | cepA in pBR322; Apr | [77] |
pHarComp | Recombination cassette for the insertion of har into P. gingivalis ECR455, cloned via A-overhangs into pGEM-T Easy; Apr | This study |
Apr, ampicillin resistant; Emr, erythromycin resistant.