T-B18
(Bacterial strain #159446)

• Purpose: E. coli BW25113 ΔfrmA, harboring plasmid Addgene plasmid # 129104, was adaptively evolved to grow efficiently on threonine and perform significant biosynthesis while employing methanol-derived carbon
• Depositing Labs: Maciek R. Antoniewicz, Eleftherios Terry Papoutsakis
• Publication: Har et al Appl Microbiol Biotechnol. 2021 Jan;105(2):869-876. doi: 10.1007/s00253-020-11058-0. Epub 2021 Jan 6.

Backbone

• Vector backbone: pETM6_Ptrc_BsMdh_BmHps_BmPhi
• Backbone manufacturer: Eleftherios Terry Papoutsakis (Addgene plasmid # 129104)
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): T-B18
• Growth instructions: LB medium with Kanamycin 25ug/mL (for ΔfrmA knockout and insertion of kanamycin resistance); Carbenicillin 100 ug/mL (vector)
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: BsMDH BmHPS BmPHI (on Addgene plasmid # 129104)
• Species: Bacillus subtilis and Bacillus methanolicus (for plasmid gene insert)
• Mutation: see Supplemental Documents for genome analysis of strain T-B18 against a reference BW25113 genome

Resource Information

• Supplemental Documents:
  • T_B18 mutations vs BW25113

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

This T-B18 E.coli strain harbors the pETM6_Ptrc_BsMdh_BmHps_BmPhi plasmid (Addgene plasmid # 129104), which enables the evolved E. coli strain to metabolize methanol in its central carbon metabolism. It was adaptively evolved from the parent strain, E. coli BW25113 ΔfrmA::kanR (obtained from the Keio collection) containing Addgene plasmid # 129104 such that it grows efficiently with just 2mM Thr and 250 mM MeOH.

This strain has been functionally validated for growth by the depositing laboratory. Addgene has not confirmed the plasmid by QC sequence analysis.