Strains |
|
|
E. coli
|
|
|
JM109 |
recA1 supE44 endA1 hsdR7 gyrA96 relA1 thi |
21 |
BL21(DE3) |
F−
ompT hsdSB(rB− mB−) dcm gal (DE3) |
Novagen |
H. mediterranei
|
|
|
DF50 |
pyrF deletion mutant of H. mediterranei ATCC 33500 |
22 |
ΔphaP mutant |
phaP deletion mutant of H. mediterranei DF50 |
20 |
ΔphaR mutant |
phaR deletion mutant of H. mediterranei DF50 |
This study |
ΔphaRP mutant |
phaRP deletion mutant of H. mediterranei DF50 |
This study |
ΔphaEC mutant |
phaEC deletion mutant of H. mediterranei DF50 |
This study |
ΔphaPEC mutant |
phaPEC deletion mutant of H. mediterranei DF50 |
This study |
ΔphaRPEC mutant |
phaRPEC deletion mutant of H. mediterranei DF50 |
This study |
E24A |
DF50 strain with PhaR carrying E24A mutation |
This study |
Q28A |
DF50 strain with PhaR carrying Q28A mutation |
This study |
Q30A |
DF50 strain with PhaR carrying Q30A mutation |
This study |
R32A |
DF50 strain with PhaR carrying R32A mutation |
This study |
K67A |
DF50 strain with PhaR carrying K67A mutation |
This study |
R75A |
DF50 strain with PhaR carrying R75A mutation |
This study |
E82A |
DF50 strain with PhaR carrying E82A mutation |
This study |
R83A |
DF50 strain with PhaR carrying R83A mutation |
This study |
Plasmids |
|
pHFX |
4.0-kb integration vector containing pyrF and its native promoter, Ampr
|
22 |
pWL502 |
7.8-kb expression vector containing pyrF and its native promoter, Ampr
|
20 |
pSCM307 |
8.2-kb shuttle vector containing promoter of hsp5 of Halobacterium sp. strain NRC-1, Ampr
|
26 |
pJAM1020 |
10.7-kb expression plasmid containing smRSGFP gene, Ampr
|
24 |
pM1915 |
8.8-kb expression vector pWL502 containing smRSGFP gene and mutated promoter of PTS |
28 |
pGEM-T Easy |
3.0-kb cloning vector, Ampr
|
Promega |
pET-28a |
5.4-kb IPTG-inducible expression vector with His6 tag |
Novagen |
pDR |
5.6-kb integration vector of pHFX for knockout of phaR
|
This study |
pDRP |
5.6-kb integration vector of pHFX for knockout of phaRP
|
This study |
pDEC |
5.3-kb integration vector of pHFX for knockout of phaEC
|
This study |
pDPEC |
5.4-kb integration vector of pHFX for knockout of phaPEC
|
This study |
pDRPEC |
5.4-kb integration vector of pHFX for knockout of phaRPEC
|
This study |
pR-IN |
5.2-kb integration vector of pHFX for knock-in of phaR
|
This study |
pRF |
8.7-kb expression vector pWL502 containing smRSGFP gene and the phaRP promoter (−151 to +17) |
This study |
pEF |
8.7-kb expression vector pWL502 containing smRSGFP gene and phaEC promoter |
This study |
pWLR |
8.4-kb expression vector pWL502 containing phaR and phaRP promoter |
This study |
pWLP |
8.5-kb expression vector pWL502 containing phaP and phaRP promoter |
20 |
pWLRP |
8.8-kb expression vector pWL502 containing phaRP and phaRP promoter |
This study |
pHP |
8.4-kb expression vector pWL502 containing phaP and hsp5 promoter |
This study |
pHRRF |
9.2-kb pRF-derived vector for additional expression of phaR under hsp5 promoter |
This study |
pRmyc |
8.3-kb pWL502 derived vector, expressing phaR-myc under mutated PTS promoter from pM1915 |
This study |
pT-Rpro |
2.9-kb pGEM-T Easy-derived cloning vector of phaRP promoter |
This study |
pM1 to pM16 |
8.7-kb pRF-derived vectors with mutations introduced into phaRP promoter |
This study |
pD41 |
8.6-kb pRF-derived vector with phaRP promoter truncated (−41 to +17) |
This study |
pD86 |
8.6-kb pRF-derived vector with phaRP promoter truncated (−86 to +17) |
This study |
pET-28aR |
5.7-kb pET-28a-derived vector, expressing phaR-His6
|
This study |
pET-28aP |
5.8-kb pET-28a derived vector, expressing phaP-His6
|
This study |
pR-IN-E24A |
5.2-kb integration vector of pHFX for knock-in of phaR with E24A mutation |
This study |
pR-IN-Q28A |
5.2-kb integration vector of pHFX for knock-in of phaR with Q28A mutation |
This study |
pR-IN-Q30A |
5.2-kb integration vector of pHFX for knock-in of phaR with Q30A mutation |
This study |
pR-IN-R32A |
5.2-kb integration vector of pHFX for knock-in of phaR with R32A mutation |
This study |
pR-IN-K67A |
5.2-kb integration vector of pHFX for knock-in of phaR with K67A mutation |
This study |
pR-IN-R75A |
5.2-kb integration vector of pHFX for knock-in of phaR with R75A mutation |
This study |
pR-IN-E82A |
5.2-kb integration vector of pHFX for knock-in of phaR with E82A mutation |
This study |
pR-IN-R83A |
5.2-kb integration vector of pHFX for knock-in of phaR with R83A mutation |
This study |