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URL: http://www.addgene.org/229395
Proper Citation: RRID:Addgene_229395
Insert Name: ΔdapA:RK2-ΔoriT:
Organism: Other
Bacterial Resistance: Other
Defining Citation: PMID:
Vector Backbone Description: Vector Backbone:N/A; Vector Types:Bacterial Expression; Bacterial Resistance:Other
Comments: The DH10B-MOB strain is a conjugation donor used to deliver plasmids containing an IncP (e.g., RK2/RP4) conjugation origin of transfer (oriT) to recipient cells. Conjugation has been used to easily deliver plasmids to a diverse range of bacteria, as well as to yeast, other fungi, and diatoms. For more information on strain construction, usage, and conjugation protocols, please see the supplementary information in “XanthoMoClo─A Robust Modular Cloning Genetic Toolkit for the Genera Xanthobacter and Roseixanthobacter.” (doi: 10.1021/acssynbio.4c00806) The strain was made using a derivative of pTA-Mob (PMCID: PMC6829330; PMC3940858). A DH10B background was made that contains this RK2/RP4 conjugation machinery—but not the oriT sequence—integrated at the dapA locus. The dapA gene is disabled, therefore, to grow the strain one must supplement growth media with diaminopimelic acid (60 ug/mL). Likewise, to select against the conjugation donor after conjugation, do not add diaminopimelic acid to the media (i.e., final recipient strain selection plates). Any plasmid containing the RK2/RP4 IncP oriT can be mobilized by this strain (AddGene #165983, AddGene #229357, AddGene #229388) and an oriT must be included on your plasmid of choice to deliver by conjugation.
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Source: Addgene
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