71431  Origami™ 2 Competent Cell Set*

Origami™ 2 host strains are K-12 derivatives that have mutations in both the thioredoxin reductase (trxB) and glutathione reductase (gor) genes, which greatly enhance disulfide bond formation in the E. coli cytoplasm. The Origami 2 strains are kanamycin sensitive; like the original Origami strains, the gor mutation is still selected for by tetracycline. To reduce the possibility of disulfide bond formation between molecules, strains containing mutations in trxB and gor are recommended only for the expression of proteins that require disulfide bond formation for proper folding.

DE3 indicates that the host is a lysogen of λDE3, and therefore carries a chromosomal copy of the T7 RNA polymerase gene under control of the lacUV5 promoter. Such strains are suitable for production of protein from target genes cloned in pET vectors by induction with IPTG.

pLysS strains express T7 lysozyme, which further suppresses basal expression of T7 RNA polymerase prior to induction, thus stabilizing pET recombinants encoding target proteins that affect cell growth and viability. Genotypes:
Origami 2
Δ(ara-leu)7697 ΔlacX74 ΔphoA PvuII phoR araD139 ahpC galE galK rpsL F′[lac⁺ lacI^q pro] gor522::Tn10 trxB (Str^R, Tet^R)

Origami 2(DE3)
Δ(ara-leu)7697 ΔlacX74 ΔphoA PvuII phoR araD139 ahpC galE galK rpsL F′[lac⁺ lacI^q pro] (DE3) gor522::Tn10 trxB (Str^R, Tet^R)

Origami 2(DE3)pLysS
Genotype: Δ(ara-leu)7697 ΔlacX74 ΔphoA PvuII phoR araD139 ahpC galE galK rpsL F′[lac⁺ lacI^q pro] (DE3) gor522::Tn10 trxB pLysS (Cam^R, Str^R, Tet^R)