E. coli strain BW25141 (pir+)

This E. coli strain can be used to propagate plasmids with R6K conditional replication origin, such as integrative base vector after the excision of its insert.

BW25141 propagates R6K origin at a copy number comparable to medium copy plasmids (~15 plasmids per cell).

CGSC Strain#:7635

Strain Designation:BW25141     Source of Strain:B.L. Wanner
Sex:F-Chromosomal Markers:Δ(araD-araB)567,ΔlacZ4787(::rrnB-3),Δ(phoB-phoR)580,λ^-,galU95,ΔuidA3::pir⁺,recA1,endA9(del-ins)::FRT,rph-1,Δ(rhaD-rhaB)568,hsdR514

Strain Comments:

• Δ(araD-araB)567-- This deletion     extends from ~25 bp upstream of the araB start codon to ~8 bp into the     beginning of the araD gene

• ΔlacZ4787(::rrnB-3)-- : 4     tandem copies of therrnBtranscriptional terminator inserted by gene replacement into the     region extending from near the SacII site near the N-terminus oflacZthrough the promoter.

• ΔuidA3::pir⁺-- Allows the     replication of plasmids with an R6Kγ origin of replication

• recA1-- : Missense     mutation, altered isoelectric point. Sequenced: G to A for Nuc. 720 (Gly     160 Asp).

• rph-1-- is a 1 bp     deletion that results in frameshift over last 15 codons and has polar     effect onpyrEleading to suboptimal pyrimidine levels on minimal medium.(Jensen     1993 JBact.175:3401)

• Δ(araD-araB)567was formerly calledΔ araBAD_AH33

• Δ(rhaD-rhaB)568was formerly calledΔ rhaBAD_LD78

• endA9(del-ins)::FRTwas formerly calledendA_BT333

• ΔlacZ4787(::rrnB-3)was formerly calledΔ lacZ_WJ16

• ΔlacZ4787(::rrnB-3)was formerly called::rrnB-4

• ΔuidA3::pir⁺was formerly calleduidA(Δ MluI)::pir⁺

• ::rrnB-3 = 4 copies of rrnB inserted

• del-ins = deletion-insertion

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References:

• Datsenko, KA, BL Wanner 2000. One-step     inactivation of chromosomal genes in Escherichia coli K-12 using PCR     products. Proc. Natl. Acad. Sci. U.S.A. 97(12):6640-5.

• Haldimann, A, BL Wanner 2001.     Conditional-replication, integration, excision, and retrieval plasmid-host     systems for gene structure-function studies of bacteria. J. Bacteriol.     183(21):6384-93.