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URL: http://www.addgene.org/61908
Proper Citation: RRID:Addgene_61908
Insert Name: none
Bacterial Resistance: Kanamycin
Defining Citation: PMID:21925267
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Comments: Genotype = cyo::kan++ ilvE avtA (Kan cassette in the chromosomal DNA) Precursor strain = ML2 (Genotype cyo::kan) Selective amino acid labeling (and/or requirement) = Ile, Leu*, Val CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR. In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes. Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system. Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
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Source: Addgene
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