pλPR cI-Elysis
(Plasmid #176891)

• Purpose: Bacterial expression plasmids for production of E. coli bacterial ghost (BG) capsules
• Depositing Lab: James Inglese
• Publication: Dranchak et al Dis Model Mech. 2023 Mar 1;16(3):dmm049863. doi: 10.1242/dmm.049863. Epub 2023 Mar 20.

Backbone

• Vector backbone: pGEM T easy
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 3015
• Total vector size (bp): 4171
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 50 μg/mL
• Growth Temperature: Room Temperature
• Growth Strain(s): DH5alpha
• Growth instructions: Growth temperature: 28C, shift to 42C for Lysis gene E induction
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Lysis E gene from PhiX174 and lambda PR-cI857 regulatory system
• Alt name: ƛPR-cI- ψX174 lysis E
• Species: PhiX174
• Insert Size (bp): 1173
• GenBank ID: KU646488
• Promoter: T7

Cloning Information

• Cloning method: TOPO Cloning
• 5′ sequencing primer: T7 promotor primer #69348-3
• 3′ sequencing primer: SP6

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: We needed to re-clone (described in comments below) based on SR Kwon et al. (2005) "Generation of Edwardsiella tarda ghosts by bacteriophage PhiX174 lysis gene E." Aquaculture 250 16-21. doi.org/10.1016/j.aquaculture.2005.02.052

Terms and Licenses

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

Depositor Comments

Lysis plasmid pλPR cI-Elysis was prepared as described previously (Kwon et al., 2005), with some modifications. PhiX174 double-stranded DNA was purchase from New England BioLabs Inc., USA (N3023S), and Lysis E gene was obtained by PCR. It is important to note that Lysis E gene on PhiX174 from New England BioLabs is mutated on two sites (587 G->A, 833 G->A), with the first mutation resulting in a premature stop codon in position 7. To override the mutations, we had to design primers that were longer than the ones described elsewhere (Kwon et al., 2005). Primer Egene-F was 7 nucleotides longer (5′-ATGGTACGCTGGACTTTGTGGGATACC-3′) and primer Egene-R was 3 nucleotides longer (5′-ACATTACATCACTCCTTCCGCAC-3′).

Please visit https://doi.org/10.1101/2022.08.26.505462 for bioRxiv preprint.