E. coli strain BW25141(λDE3)

BioVector NTCC質(zhì)粒載體菌種細(xì)胞蛋白抗體基因保藏中心www.biovector.net

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.