Acinetobacter ADP1

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Cultures

  • ADP1 can be grown anywhere between room temperature and 37°C
  • Will not survive at 4°C
  • For long term storage, add 10% DMSO and store at -80°C
  • Naturally resistant to ampicillin and chloramphenicol. Use spectinomycin, kanamycin, or tetracycline for selections.

Transformations

(derived from Metzgar et al. paper)

  • Streak out cells on LB plate, grow overnight at 37°C
  • Innoculate colonies into 2 mL LB and grow for 12-16 hours at 30°C with shaking
  • Take a 20 μL aliquot and add to 280 μL fresh LB
  • Add DNA
  • Shake at 30°C for 2 hours
  • Plate on appropriate media with antibiotic and let grow at 30°C

Constructs

  • 2.1 in plasmid IMBB
    • IMBB expresses TEM-1 β-lactamase and has a ColE1 origin
  • 2.2 in plasmid IMBB
  • 2.1-P1010-2.2 in plasmid pSB1AC3
  • 2.1-J04450-P1010-2.2 in plasmid pSB1AC3
  • gusA-specR reporter system
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