Escherichia coli DH10B-MOB
(Bacterial strain #229395)

• Purpose: Escherichia coli DH10B DAP-auxotrophic strain containing an integrated RK2/RP4 mobilization vector to conjugate plasmids with an IncP origin of transfer (modified pTA-Mob 2.0, AddGene Plasmid #149662)
• Depositing Lab: Michael Springer
• Publication: Soltysiak et al ACS Synthetic Biology. 2025. doi: 10.1021/acssynbio.4c00806.

Backbone

• Vector backbone: N/A
• Vector type: Bacterial Expression
• Selectable markers: Diaminopimelic Acid (DAP) Auxotroph

Growth in Bacteria

• Bacterial Resistance(s): 60 ug/mL of DAP
• Growth Temperature: 37°C
• Growth Strain(s): DH10B-MOB
• Growth instructions: Requires 60 ug/mL of DAP
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: ΔdapA:RK2-ΔoriT:
• Species: Escherichia coli

Cloning Information

• Cloning method: Unknown

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.