KJ740(DE3)
(Bacterial strain #179486)

• Purpose: KJ740(DE3) is an OmpC-, OmpF-deletion strain of E. coli that allows for T7 promoter-based expression
• Depositing Lab: Debra Hansen
• Publication: Boyd et al FASEB J. 2022 Jul;36(7):e22378. doi: 10.1096/fj.202101808RR.

Backbone

• Vector backbone: N/A

Growth in Bacteria

• Bacterial Resistance(s): Tetracycline (chromosome, Tn10) (10 μg/mL); Streptomycin/Spectinomycin (chromosome) (50 μg/mL)
• Growth Temperature: 37°C
• Growth Strain(s): N/A
• Growth instructions: This strain can be grown in LB Miller medium (10 g tryptone, 5 g yeast extract, and 10 g sodium chloride per liter)
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: none
• Species: E.coli

Resource Information

• Supplemental Documents:
  • PCR validation of λ(DE3) lysogen in strain KJ740(DE3)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

λDE3 [lacI, lacUV5-T7 gene 1, ind1, sam7, nin5] F- araC14 fhuA31 lacY1 tsx-78 ΔompF480 zcb-222::Tn10 rfbC1 mgl-51 ΔompC178 rpsL136(StrR) xylA5 mtl-1 thiE1

KJ740(DE3) is a λDE3 derivative of the OmpC- & OmpF-deficient strain KJ740 that was described in Ingham C, Buechner M, Adler J, 1990, J Bacteriol 172:3577-3583.

Due to the presence of the lambda (DE3) lysogen, KJ740(DE3) is compatible with expression from the T7 promoter.

The (DE3) lysogen of the parent strain KJ740 was made by Debra T. Hansen using the λDE3 Lysogenization Kit 538 (EMD Millipore #69734-3). The parent strain KJ740 was obtained from the Yale E. coli Genetic Stock Center (CGSC) as strain number 12151.

In contrast to BL21 derivative strains, KJ740(DE3) is not deficient in the proteases Lon and OmpT.

The presence of (DE3) was confirmed by PCR by Felicia M. Craciunescu.

PCR primers that may be used to verify the presence of the (DE3) lysogen:
These primer pairs are expected to generate a PCR product from each of 3 genes within the (DE3) sequence, GenBank accession no. AM946981.2. There are two primer pairs per gene, therefore 6 possible PCR products. The presence of the PCR product of the expected size indicates the presence of the (DE3) lysogen. The PCR reactions will also be run on the negative control non-(DE3) parent strain, KJ740, which is expected to not generate these PCR product sizes.

gene A F1 GACTACATCCGTGAGGTGAATGTGG
gene A R1 GATGACGCAGGCATTATGCTCGCAG
Product size = 645 bp

gene A F2 CCTTTCACATCTGGACAGCGTACAG
gene A R2 GTTGCTGCACCATCCTCTTCCTGC
Product size = 904 bp

gene H F1 GACATCTGGAATCTGCGCAAGGATGA
gene H R1 CTGCAGGATTTTCCCGTCTTTCAGTG
Product size = 291 bp

gene H F2 GTGCGCTGGCGTATGCCTGGTATC
gene H R2 CTGTTCCGTGGCTTCCCGTTCTGC
Product size = 1388 bp

gene J F1 CTTTACCTTCGGTGTACAGGCACTGGTG
gene J R1 GTCCTGCACGAACGTCAGCGTCTG
Product size = 748 bp

gene J F2 CAGACAGGTTGAAACCAGCACGCGTTATC
gene J R2 GTTTGCCTTCCTCCGTGTCCTCCATG
Product size = 407 bp

Please visit https://www.biorxiv.org/content/10.1101/2021.08.26.457821v2 for bioRxiv preprint.