Lysate catalog
In the following we list the characteristics of the cell lysates we produced in our labaratory for the purposes of improving cell lysates characterization and finding the most suited lysates for our detection tool.
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BL21(DE3) Lysate
- Organism: E. coli
- Strain: BL21(DE3)
- Bacterial strain supplier: Novagen1
- Genotype: F-ompT hsdSB(rB-mB-) gal dcm (DE3)
- Polymerase(s):
- E.coli RNA polymerase
- T7 RNA polymerase (gene under control of the lacUV5 promoter induced by IPTG)
- Antibiotic Resistances:
- None
- Comments:
- Engineered for transformation and high protein expression
- Deficient in Ion and ompT protease
- T7 RNA polymerase expression when induced with IPTG
M15 Lysate
- Organism: E. coli
- Strain: E. coli M15 DZ291
- Bacterial strain supplier: Qiagen2
- Genotype: F- Φ80ΔlacM15 thi lac- mtl- recA+KmR
- Polymerase(s):
- E. coli RNA polymerase
- Antibiotic Resistances:
- Kanamycin
- Comments:
- Deficiency in lacZα
- blue-white screening or complementation of β-galactosidase
T7-M15 Lysate
- Same characteristics as M15 strain
- M15 strain transformed with plasmid pQE30 with the T7 RNA polymerase gene inserted between BamH1 and HindIII restriction sites3
- Polymerase(s):
- E. coli RNA polymerase
- T7 RNA polymerase
- Antibiotic Resistance:
- Ampicillin
- Kanamycin
- Comments:
- T7 RNA polymerase expression when induced with IPTG
DH5α
- Organism: E. coli
- From: Thermofisher4
- Strain: DH5α™ T1R
- Genotype: F– Φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK–, mK+) phoA supE44 thi-1 gyrA96 relA1 tonA
- Polymerase(s):
- E. coli RNA polymerase
- Resistances:
- None
- Comments:
- Resistance to T1 and T5 phage
- Deficiency in lacZα
- blue-white screening or complementation of β-galactosidase
Top10
- Organism: E. coli
- From: Invitrogen5
- Strain: TOP10
- Genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 Δ lacX74 recA1 araD139 Δ(araleu)7697 galU galK rpsL (StrR) endA1 nupG
- Polymerase(s):
- E. coli RNA polymerase
- Resistances:
- None
- Comments:
- Deficiency in lacZα
- blue-white screening or complementation of β-galactosidase
Top10-gams
- Organism: E. coli
- From: Invitrogen 6
- Strain: TOP10
- Genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 Δ lacX74 recA1 araD139 Δ( araleu)7697 galU galK rpsL (StrR) endA1 nupG
- Polymerase(s):
- E. coli RNA polymerase
- Resistances:
- Ampicillin
- Comments:
- Deficiency in lacZα
- blue-white screening or complementation of β-galactosidase
- Plasmid pBADmod1-linker2-gamS7 is inserted in top10 competent cells to produce gamS
- Respectively T7 RNA polymerase and gamS protein expression when induced with IPTG and arabinose
