TABLE 1.
Bacterial strains and plasmids
Strain or plasmid | Description | Source or reference |
---|---|---|
Strains | ||
E. coli K-12 | Wild type | DSMZ |
C41(DE3) | Derived from BL21 [F−ompT hsdSB(rB− mB−) gal dcm (DE3)]; has at least one uncharacterized mutation that prevents cell death associated with expression of many toxic recombinant proteins | 15 |
R721 | 71/18 glp O-P434/p22lacZ | 5 |
Plasmids | ||
pET22b(+) | N-terminal pelB signal sequence plus an optional C-terminal His-tagged sequence; ampicillin resistant | Novagen |
pET28a(+) | N-terminal His tag-thrombin-T7 tag configuration plus optional C-terminal His tag sequence; kanamycin resistance | Novagen |
pET22b-αpol | pET22b(+) Δ(NdeI-XhoI) Ω(rpoA) | This study |
pET22b-c-Myc | pET22b(+) Δ(NdeI-BamHI) Ω(c-myc tag) | This study |
pET22b-c-Myc-L2 | pET22b-c-Myc Δ(BamHI-XhoI) Ω(rplB) | This study |
pET22b-c-Myc-Thioredoxin | pET22 b-c-Myc Δ(BamHI-XhoI) Ω(trxC) | This study |
pET28a-αpol | pET28a Δ(BamHI-XhoI) Ω(rpoA) | This study |
pcIP22 | pC132 derivative carrying N-terminal end of p22 repressor; ampicillin resistance | 5 |
pcI434 | pACYC177 derivative carrying N-terminal end of 434 repressor; chloramphenicol resistance | 5 |
pcIP22-RNAPα | pcIP22Δ(SalI-BamHI) Ω(rpoA) | This study |
pcI434-RNAPa | pcI434Δ(SalI-BamHI) Ω(rpoA) | This study |
pcI434-RPL2 | pcI434Δ(SalI-BamHI) Ω(rplB) | This study |
pcI-L2 | pcI434 Δ(NdelI-BamHI) Ω(rplB) | This study |
pcI-L1 | pcI434Δ(NdelI-BamHI) Ω(rplA) | This study |
pcI-L3 | pcI434Δ(NdelI-BamHI) Ω(rplC) | This study |
pcI-L20 | pcI434Δ(NdelI-BamHI) Ω(rplT) | This study |
pcI-L27 | pcI434Δ(NdelI-BamHI) Ω(rpmA) | This study |
pET22b-PrrnD | pET22b Δ(SphI-HindIII) Ω(rrsD promoter) | This study |
pET22b-PaidB | pET22b Δ(SphI-HindIII) Ω(aidB promoter) | This study |
pET22b-PrrnD-lacZ | pET22b-PrrnDΔ(HindIII-XhoI) Ω(lacZ) | This study |
pET22b-PaidB-lacZ | pET22b-PaidBΔ(HindIII-XhoI) Ω(lacZ) | This study |