MG1655/pKD46      Source of Strain:
pKD46 Plasmid Markers/Mutations: repA101
, [tL3] Plasmid Comment:
Lambda Red recombinase expression plasmid -- includes 2154 nucleotides (31088-33241) of phage Lambda, which includes the terminator downstream frp, exo, tL3 and a small ORF.GenBk-AY048746 Chromosomal Markers: λ-
, rph-1Strain Comments:
- pKD46 is more efficient than pKD20 for making gene disruptions. pKD46 sequence: GenBank AY048746
- The pKD46 plasmid seems to be less temperature sensitive when in this host. We are currently investigating this phenomenon.
- This strain should be grown on rich media with ampicillin (100 ug/ml) at 30 C.
- rph-1-- is a 1 bp deletion that results in frameshift over last 15 codons and has polar effect on pyrE leading to suboptimal pyrimidine levels on minimal medium.(Jensen 1993 JBact.175:3401)
- ApR = ampicillin resistant
- ts = Temperature Sensitive
- 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.