TABLE 1.
A. baylyi strains and plasmids
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| A. baylyi strains | ||
| ADP1 | Wild type (BD413) | 18 |
| ISA36 | benM::ΩS4036 | 12 |
| ADP102 | catM3102b | 23 |
| ACN47 | benM::ΩS4036 benA::lacZ-Kmr5032 | 12 |
| ACN146 | benM::ΩS4036 benMA5146 | 12 |
| ACN153 | benM::ΩS4036 catM5153 [CatM(V158M)] | This study |
| ACN157 | benA::lacZ-Kmr5032 benM::ΩS4036 benMA5146 | 12 |
| ACN164 | benM::ΩS4036 benA::lacZ-Kmr5032 catM5153 [CatM(V158M)] | This study |
| ACN293 | benM::ΩS4036 benMA5147 ΔcatM5293 | 27 |
| ACN539 | benM::ΩS4036 benMA5147 ΔcatM5293 catB::lacZ-Kmr5534 | This study |
| ACN541 | benM::ΩS4036 benMA5146 catMΩK5541 | This study |
| ACN547 | benM::ΩS4036 benMA5146 catM3102 [CatM(R156H)] | This study |
| ACN548 | benM::ΩS4036 benMA5146 benA::lacZ-Kmr5032 catM3102 [CatM(R156H)] | This study |
| ACN549 | catM3102 [CatM(R156H)] | This study |
| ACN558 | benM::ΩS4036 catM3102 [CatM(R156H)] | This study |
| ACN559 | benM::ΩS4036 benA::lacZ-Kmr5032 catM3102 [CatM(R156H)] | This study |
| ACN560 | benM::ΩS4036 catB::lacZ-Kmr5534 catM5153 [CatM(V158M)] | This study |
| ACN561 | benM::ΩS4036 benMA5146 catB::lacZ-Kmr5534 catM3102 [CatM(R156H)] | This study |
| ACN585 | benM::ΩS4036 catB::lacZ-Kmr5534 | This study |
| Plasmids | ||
| PUC13, pUC19 | Apr; cloning vector | 36 |
| pRK415 | Tcr; broad-host-range cloning vector | 19 |
| pCR2.1-TOPO | Apr; PCR cloning vector | Invitrogen |
| pET-21b | Apr; T7 expression vector | Novagen |
| pHP45 | Apr Smr Spr; source of ΩS | 25 |
| pKOK6 | Apr Kmr; source of promoterless lacZ-Kmr cassette | 20 |
| pUI1637 | Apr Kmr; source of ΩK | 14 |
| pIB17 | Apr; catM3102 (11950-13205)c in pUC19 | 23 |
| PIGG4 | Apr; catM fragment (11950-12892)c in pUC19 | This study |
| pIGG14 | Apr Smr Spr; catBCIJFD (12892-18153)c in pUC13 with ΩS inserted in EcoRV site (15660)c of catJ | This study |
| pBAC6 | Apr Kmr; fragment (11950-12892)c containing catM in pUC19 (ΩK in HincII site [12687]c of catM) | This study |
| pBAC44 | Apr Smr Spr; benMABCDE (563-7876)c in pUC19 (ΩS in multiple cloning site of vector) | This study |
| pBAC54 | Apr Kmr; lacZ-Kmr cassette in NsiI site (3761)c in benA with adjacent ben region (2316-5663)c in pUC19 | 12 |
| pBAC200 | Tcr; cat region DNA (9819-10649 and 15951-18153)c in pRK415; used to isolate the chromosomal catMB region | 13 |
| pBAC234 | Tcr; ACN153 cat region DNA (9819-18153)c in pRK415; isolated with pBAC200, catM5153 mutation | This study |
| pBAC284 | Apr; catM segment (11950-12892)c from pBAC234 cloned in pUC19, catM5153 mutation | This study |
| pBAC364 | Apr; benMA5146 intergenic region (2293-2540)c,d in pUC19, DNase I footprinting, antisense strand labeling | This study |
| pBAC366 | Apr; wild-type benMA intergenic region (2293-2540)c,d in pUC19, DNase I footprinting, antisense strand labeling | 6 |
| pBAC371 | Apr; benMA5146 region (2316-2540),c,d HindIII deletion of pBAC364, DNase I footprinting, sense strand labeling | This study |
| pBAC373 | Apr; wild-type benMA region (2316-2540),c,d HindIII deletion of pBAC366, DNase I footprinting, sense strand labeling | 6 |
| pBAC383 | Apr; PCR fragment with catM5153 in pET-21b, for purification of CatM(V158M) | This study |
| pBAC669 | Apr; catBCIJ (12892-15658)c in pUC13 | This study |
| pBAC673 | Apr; catB (13205-14225)c in pUC19 | This study |
| pBAC674 | Apr Kmr; catB (13205-14225)c upstream of lacZ-Kmr cassette in SalI site (14225)c of catB in pUC19 | This study |
| pBAC675 | Apr Kmr; catB (13205-14225)clacZ-KmrcatJFD (15660-17347)c in pUC19 | This study |
| pBAC679 | Apr Kmr; PCR fragment with catM3102 in pCR2.1-TOPO | This study |
| pBAC684 | Apr; 0.91-kb catM3102 NdeI-XhoId fragment from pBAC679 in pET-21b, complete coding sequence | This study |
a
Apr, ampicillin resistant; Tcr, tetracycline resistant; Smr, streptomycin resistant; Spr, spectinomycin resistant; Kmr, kanamycin resistant; ΩS, omega cassette conferring Smr Spr; ΩK, omega cassette conferring Kmr.
b
The original strain isolated with the catM3102 allele has a mucoid colony morphology and is likely to carry additional uncharacterized mutations.
c
Position in the ben-cat sequence in GenBank entry (accession number AF009224).
d
Restriction sites added in primers used to generate PCR-amplified fragment for cloning.