E. coli strain BW25141(λDE3)

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The E. coli BW25141strain can be used to propagate plasmids with R6K conditional replicationorigin, such as integrative base vector after the excision of its insert.

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

Strain Designation:BW25141    Source of Strain:B.L. WannerSex: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

StrainComments:

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

·ΔlacZ4787(::rrnB-3)--: 4     tandem copies of the rrnB transcriptional terminator inserted by gene replacementinto the     region extending from near the SacII site near theN-terminus of lacZ through the promoter.

·ΔuidA3::pir⁺-- Allows the     replication of plasmids withan R6Kγ origin of replication

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

·rph-1-- isa 1 bp     deletion that results in frameshift over last 15 codonsand has polar     effect onpyrEleading to suboptimal pyrimidine levels on minimalmedium.(Jensen     1993 JBact.175:3401)

·Δ(araD-araB)567was formerly called Δ araBAD_AH33

·Δ(rhaD-rhaB)568was formerly called Δ rhaBAD_LD78

·endA9(del-ins)::FRTwas formerly called endA_BT333

·ΔlacZ4787(::rrnB-3)wasformerly called Δ lacZ_WJ16

·ΔlacZ4787(::rrnB-3)wasformerly called::rrnB-4

·ΔuidA3::pir⁺was formerly called uidA(Δ MluI)::pir⁺

·::rrnB-3= 4 copies of rrnB inserted

·del-ins= deletion-insertion

·

References:

·Datsenko,KA, BL Wanner 2000. One-step     inactivation of chromosomal genes inEscherichia 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 genestructure-function studies of bacteria. J. Bacteriol.    183(21):6384-93.