Acinetobacter ADP1

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