Strains |
|
|
YMC10 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs
|
8 |
RB9060 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306
|
10 |
K |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs Δmdl-glnK::Kan |
6 |
BK |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::Kanr
|
6 |
BKc |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::Camr
|
6 |
BKΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
6 |
WCH30 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnK1 (amtB+) |
1 |
UNF3435 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnK1 (amtB+) ΔglnB2306
|
1 |
YMC15 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302
|
11 |
BKgΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnK1 (amtB+) ΔglnB2306 trpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
BΦ × WCH30 P1vir
|
BΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 trpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
6 |
BA |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 glnA::Tn5
|
13 |
TEΦ |
recD1903::Tn10 trpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
6 |
YMC10Φ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs trpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
6 |
YMC15Φ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 trpDC700::putPA130 [Φ(glnKp-lacZ) Kanr Cams] |
YMC15 × TEΦ P1vir
|
MAKc |
recD Δmdl-glnK::Camr
|
6 |
Kc |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs Δmdl-glnK::Camr
|
YMC10 × MAKc P1vir
|
YMC15K |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::Camr
|
YMC15 × MAKc P1vir
|
YMC15B |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306
|
BA × YMC15 P1vir
|
YMC15BΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306 trpDC700::putPA1303 [Φ(glnKp-lacZ) Kanr Cams] |
YMC15B × TEΦ P1vir
|
YMC15BKΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Φ(glnKp-lacZ) Kanr Cams] |
YMC15BΦ × MAKc P1vir
|
YMC15KΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::CamrtrpDC700::putPA1303 [Φ(glnKp-lacZ) Kanr Cams] |
YMC15Φ × MAKc P1vir
|
Y15KgΦ |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnK1 (amtB+) trpDC700::putPA1303 [Φ(glnKp-lacZ) Kanr Cams] |
YMC15Φ × WCH30 P1vir
|
TE2680 |
recD1903::Tn10 trpDC700::putPA1303 [Kans Camrlac] |
12 |
MABpB |
recD1903::Tn10 trpDC700::putPA1303 [Kanr CamsglnB8] |
TE2680 × pglnB8 DNA |
MAKpK |
recD1903::Tn10 trpDC700::putPA1303 [Kanr CamsglnK91] |
TE2680 × pglnK91 DNA |
MAKpB2 |
recD1903::Tn10 trpDC700::putPA1303 [Kanr Cams Φ(glnKp-glnB)] |
TE2680 × pglnKpB2 DNA |
MABpK2 |
recD1903::Tn10 trpDC700::putPA1303 [Kanr Cams Φ(glnBp-glnK)] |
TE2680 × pglnBpK2 DNA |
MAApB2 |
recD1903::Tn10 trpDC700::putPA1303 [Kanr Cams Φ(glnAp-glnB)] |
TE2680 × pglnApB2 DNA |
MAApK2 |
recD1903::Tn10 trpDC700::putPA1303 [Kanr Cams Φ(glnAp-glnK)] |
TE2680 × pglnApK2 DNA |
BKcBpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr CamsglnB8] |
BKc × MABpB P1vir
|
BKcBpK2 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr Cams Φ(glnBp-glnK)] |
BKc × MABpK2 P1vir
|
BKcKpK |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr CamsglnK91] |
BKc × MAKpK P1vir
|
BKcKpB2 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr Cams Φ(glnKp-glnB)] |
BKc × MAKpB2 P1vir
|
BKcApB2 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr Cams Φ(glnAp-glnB)] |
BKc × MAApB2 P1vir
|
BKcApK2 |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs ΔglnB2306 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr Cams Φ(glnAp-glnK)] |
BKc × MAApK2 P1vir
|
Y15KcBpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr CamsglnB8] |
YMC15K × MABpB P1vir
|
Y15KcKpK |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr CamsglnK91] |
YMC15K × MAKpK P1vir
|
Y15KcKpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::CamrtrpDC700::putPA1303 [Kanr Cams Φ(glnKp-glnB)] |
YMC15K × MAKpB2 P1 vir
|
Y15BBpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306 trpDC700::putPA1303 [Kanr CamsglnB8] |
YMC15B × MABpB P1vir
|
Y15BKpK |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306 trpDC700::putPA1303 [Kanr CamsglnK91] |
YMC15B × MAKpK P1vir
|
Y15BKpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 ΔglnB2306 trpDC700::putPA1303 [Kanr Cams Φ(glnKp-glnB)] |
YMC15B × MAKpB2 P1vir
|
Y15BKcBpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::Camr ΔglnB2306 trpDC700::putPA1303 [Kanr CamsglnB8] |
Y15BBpB × MAKc P1vir
|
Y15BKcKpK |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::Camr ΔglnB2306 trpDC700::putPA1303 [Kanr CamsglnK91] |
Y15BKpK × MAKc P1vir/PICK> |
Y15BKcKpB |
endA1 thi-1 hsdR17 supE44 ΔlacU169 hutCklebs glnL2302 Δmdl-glnK::Camr ΔglnB2306 trpDC700::putPA1303 [Kamr Cams Φ(glnKp-glnB)] |
Y15BKpB × MAKc P1vir
|
Plasmids |
|
|
pRS551 |
lac fusion vector |
36 |
pAlter-1 |
Amps mutagenesis vector |
Promega Corporation |
pUC18N |
pUC18, with NdeI site destroyed |
6 |
pUC18NS |
pUC18, with NdeI and SalI sites destroyed |
|
pglnB5 |
glnB PCR product cloned as BamHI fragment into pAlter-1 |
|
pglnB6 |
NdeI site introduced at glnB start codon of pglnB5 |
|
pglnB8 |
BamHI fragment containing glnB cloned into pRS551 from pglnB6 |
|
pglnB10 |
glnB cloned into pUCN as EcoRI/HindIII fragment from pglnB6 |
|
pglnK51 |
glnK PCR product cloned as BamHI fragment into pAlter-1 |
|
pglnK71 |
NdeI site introduced at glnK start codon of pglnK5 |
|
pglnK81 |
BamHI fragment containing glnK cloned into pUCNS from pglnK71 |
|
pglnK91 |
BamHI fragment containing glnK cloned into pRS551 from pglnK71 |
|
pglnKpB1 |
glnK coding region of pglnK81 exchanged for the glnB coding region from pglnB10 as NdeI/SalI fragment |
|
pglnKpB2 |
BamHI fragment containing glnKp-glnB fusion gene cloned into pRS551 |
|
pglnBpK1 |
glnB coding region of pglnB10 exchanged for the glnK coding region of pglnK71
|
|
pglnBpK2 |
BamHI fragment containing glnBp-glnK fusion gene cloned into pRS551 |
|
pglnApB1 |
glnB promoter of pglnB10 exchanged for glnA promoter generated by PCR as a KpnI/NdeI fragment |
|
pglnApB2 |
BamHI fragment containing glnAp-glnB fusion gene cloned into pRS551 |
|
pglnApK1 |
glnB coding region of glnAp-glnB fusion gene exchanged for the glnK coding region of glnK71 as NdeI/HindIII fragment |
|
pglnApK2 |
BamHI fragment containing glnAp-glnK fusion gene cloned into pRS551 |
|
Primers |
|
|
113D |
GGGGATCCGCGGCCCGGTTGGCAACATGC, primer downstream of glnB for cloning (BamHI) |
|
7158I |
GCTGCAGGGATCCCTCAACTATTTGCGTAAGCTGCTGC, cloning primer located upstream of glnB promoter (PstI/BamHI) |
|
7159I |
GCTGCAGGGATCCCATTGAGCGCCTGAATAGCGC, cloning primer located upstream of glnk promoter (PstI/BamHI) |
|
5048F |
CGGATCCGTCGACTTCCTGTTGCTGTGTGCCAGAG, primer downstream of glnK for cloning and sequencing (SalI/BamHI) |
|
4158J |
CGGTACCGGATCCCCTCCGCAAACAAGTATTGCAGAGTCCC, cloning primer located upstream of glnA promoter (KpnI/BamHI) |
|
4157J |
CCATATGTTAACTCTCCTGGATTGGTCATGGTCGTCGTG downstream primer introduces NdeI site at glnA ATG start codon for cloning the glnA promoter region (NdeI) |
|
9829I |
GACCGGAGGGGACATATGAAGCTGGTGACC, internal primer for site-specific mutagenesis introducing an NdeI |
|
|
site at the glnK ATG start codon |
|
9251I |
TTTTCAAGGAATCATATGAAAAAGATTGAT, internal primer for site-specific mutagenesis introducing an NdeI site at the glnB ATG start codon |
|
4588E |
GGTCATCAGCAATCGCCAC, internal glnK sequencing primer |
|
5551E |
CGTCAGTAAGGCGGCTTACAC, internal glnK sequencing primer |
|
4929F |
GATTATCACGGTCACCAGCTTC, internal glnK sequencing primer |
|
2820 |
ACGGTGACCGAAGTGAAAGGC, internal glnB sequencing primer |
|
2866 |
CAGCTCGGTATGGCCTTTC, internal glnB sequencing primer |
|
3098 |
CGATCCGGCAACCCTTGACGCGG, internal glnB sequencing primer |
|