Here you will find a collection of E.coli strains we use and offer in our PPCF for members of CSSB groups. Please note that due to the existence of restricting material transfer agreements (MTAs) for some strains, those strains can only be provided for use in the Protein Production Core Facility. If you are interested in using these vectors or strains in a lab other than the PPCF, please get in contact so we can help you to put all needed MTAs in place. Some MTAs are coupled with fees. We are happy to advise you on this issue.
Cloning
| Name | Manufactor | parental | Antibiotic resistance | Genotype | remarks |
|---|---|---|---|---|---|
| One Shot™ ccdB Survival™ 2 T1R | Invitrogen | K12 | Streptomycin | F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG fhuA::IS | for propagation of ccdB cassette , not suitable for pCoofy |
| Top10 | Invitrogen | K12 | Streptomycin | F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG | for routine cloning, including mammalian and plant DNA, for methylated DNA |
| DB3.1 | Life Technologies Corporation /BCCM | K12 | Streptomycin | gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20 glnV44 (=supE44) ara14 galK2 lacY1 proA2 rpsL20 xyl5 leuB6 mtl1 | for propagation of ccdB cassette, suitable for pCoofy |
| XL10-gold | Stratagene/Agilent | K12 | Tetracycline, Chloramphenicol | endA1 glnV44 recA1 thi-1 gyrA96 relA1 lac Hte Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 tetR F'[proAB lacIqZΔM15 Tn10(TetR Amy CmR)] | Hte phenotype allows high transformation with large plasmid inserts |
| DH10Bac | Gibco/ThermoFisher | K12 | Tetracycline, Kanamycin, Gentamycin | F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ-rpsL nupG/pMON14272/pMON7124 | production of recombinant bacmids used in BVES |
Expression
| Name | Manufactor | parental | Antibiotic Resistance | Genotype | remarks |
|---|---|---|---|---|---|
| SHuffle T7 | NEB | K12 | Streptomycin | F´ lac, pro, lacIq / Δ(ara-leu)7697 araD139 fhuA2 lacZ::T7 gene1 Δ(phoA)PvuII phoR ahpC* galE (or U) galK λatt::pNEB3-r1-cDsbC (SpecR, lacIq) ΔtrxB rpsL150(StrR) Δgor Δ(malF)3 | suitable for T7 protein expression with enhanced capacity to correctly fold proteins with multiple disulfide bonds in the cytoplasm |
| SHuffle T7 express LysY | NEB | B | Chloramphenicol | MiniF lysY (CamR) / fhuA2 lacZ::T7 gene1 [lon] ompT ahpC gal λatt::pNEB3-r1-cDsbC (SpecR, lacIq) ΔtrxB sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10 --TetS) endA1 Δgor ∆(mcrC-mrr)114::IS10 | suitable for T7 protein expression with enhanced capacity to correctly fold proteins with multiple disulfide bonds in the cytoplasm and with enhanced reduction of basal expression, Control of T7 RNA Polymerase by lysozyme allows potentially toxic genes to be expressed |
| SHuffle express | NEB | B | Streptomycin | fhuA2 [lon] ompT ahpC gal λatt::pNEB3-r1-cDsbC (SpecR, lacIq) ΔtrxB sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10 --TetS) endA1 Δgor ∆(mcrC-mrr)114::IS10 | suitable for non-T7 protein expression |
| SHuffle T7 express | NEB | B | Streptomycin | fhuA2 lacZ::T7 gene1 [lon] ompT ahpC gal λatt::pNEB3-r1-cDsbC (SpecR, lacIq) ΔtrxB sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10 --TetS) endA1 Δgor ∆(mcrC-mrr)114::IS10 | suitable for T7 protein expression with enhanced capacity to correctly fold proteins with multiple disulfide bonds in the cytoplasm |
| BL21 | Merck / Novagen | B | --- | F– ompT gal dcm lon hsdSB(rB–mB–) [malB+]K-12(λS) | non-T7 expression E. coli strain, does not express the T7 RNA Polymerase, Ideal for Plac, Ptac, Ptrc ParaBAD expression vectors |
| BL21(DE3) | Merck / Novagen | B | --- | F– ompT gal dcm lon hsdSB(rB–mB–) λ(DE3 [lacI lacUV5-T7p07 ind1 sam7 nin5]) [malB+]K-12(λS) | Ideal for routine T7 expression |
| BL21(DE3) pLysS | Merck / Novagen | B | Chloramphenicol | F– ompT gal dcm lon hsdSB(rB–mB–) λ(DE3 [lacI lacUV5-T7p07 ind1 sam7 nin5]) [malB+]K-12(λS) pLysS[T7p20 orip15A](CmR) | pLysS plasmid encodes T7 phage lysozyme, an inhibitor for T7 polymerase which reduces and almost eliminates expression from transformed T7 promoter containing plasmids when not induced |
| Rosetta2(DE3) | Merck/ Novagen | B | Chloramphenicol | F- ompT hsdSB(rB- mB-) gal dcm (DE3) pRARE2 (CamR) | designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E.coli. These strains supply tRNAs for 7 rare codones (AGA, AGG, AUA, CUA, GGA, CCC, and CGG) on a compatible chloramphenicol-resistant plasmid. |
| ArcticExpress(DE3) | Agilent | B | Gentamycin | E. coli B F– ompT hsdS(rB– mB–) dcm+Tetr galλ(DE3) endA Hte [cpn10 cpn60 Gentr] | suitable for T7 protein expression, co-expression of the cold-adapted chaperonins Cpn10 and Cpn60 from the psychrophilic bacterium Oleispira antarctica. These chaperonins confer improved protein processing at lower temperatures, potentially increasing the yield of active, soluble recombinant protein. Cells are grown for 3h at 30°C without selection marker, expression is induced with IPTG, cells are grown at 10-13°C for 24h. |
| C43(DE3) | Sigma-Aldrich | --- | F – ompT hsdSB (rB- mB-) gal dcm (DE3) | The strain C43(DE3) was derived from C41(DE3) by selecting for resistance to a different toxic protein and can express a different set of toxic proteins to C41(DE3). Effective in expressing toxic & membrane proteins. | |
| Origami B(DE3) | Merck | Kanamycin, Tetracycline | F- ompT hsdSB(rB- mB-) gal dcm lacY1 ahpC (DE3) gor522:: Tn10 trxB (KanR, TetR) | carry the same mutations in trxB and gor as the original Origami strains, except that they are derived from a lacZY mutant of BL21 to enable precise control of expression levels by adjusting the concentration of IPTG. Thus the Origami B strains combine the desirable characteristics of BL21, Tuner™, and Origami strains in one strain background. The mutations in trxB and gor are selectable on kanamycin and tetracycline, respectively; therefore, these strains cannot be used with plasmids that can only selected with kanamycin or tetracycline. These strains also include the lon and ompT deficiences of BL21, which increase protein stability. | |
| Tuner(DE3) | Novagen | B | --- | F–ompT hsdSB (rB– mB–) gal dcm lacY1(DE3) | lacZY deletion mutants of BL21 and enable adjustable levels of protein expression throughout all cells in a culture. The lac permease (lacY) mutation allows uniform entry of IPTG into all cells in the population, which produces a concentration-dependent, homogeneous level of induction. By adjusting the concentration of IPTG, expression can be regulated from very low levels up to the robust, fully induced levels commonly associated with pET vectors. Lower-level expression may enhance the solubility and activity of difficult target proteins |
| Rosetta gami 2(DE3) | Novagen | Chloramphenicol | Δ(ara-leu)7697 ΔlacX74 ΔphoA PvuII phoR araD139 ahpC galE galK rpsL (DE3) F′[lac+ lacIq pro] gor522::Tn10 trxB pRARE2 (CamR, StrR, TetR) | combine the advantages of Rosetta 2 and Origami 2 strains to alleviate codon bias and enhance disulfide bond formation in the cytoplasm when heterologoous proteins are expressed in E. coli. These trxB/gor mutants are compatible with kanamycin-resistant vectors. The cells carry the chloramphenicol-resistant plasmid, pRARE2, which supplies tRNAs for seven rare codons, AUA, AGG, AGA, CUA, CCC, GGA, and CGG under the control of their native promoters. | |
| Lemo21(DE3) | NEB | Chloramphenicol | fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo(CamR) λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5 pLemo = pACYC184-PrhaBAD-lysY | Lemo21(DE3) offers the host features of BL21(DE3) while also allowing for tunable expression of difficult clones. Tunable expression is achieved by varying the level of lysozyme (lysY), the natural inhibitor of T7 RNA polymerase. The level of lysozyme is modulated by adding L-rhamnose to the expression culture at levels from zero to 2000 µM. When Lemo21(DE3) is grown without rhamnose, the strain performs the same as a pLysS containing strain. However, optional addition of rhamnose tunes the expression of the protein of interest. For difficult soluble proteins, tuning the expression level may also result in more soluble, properly folded protein. |
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