Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Antibiotic Resistance

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10−8 to 10−9) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic MIC Standard Conc. Status Notes
Ampicillin (or carbenicillin) 128 g/ml 100 g/ml Resistant  
Kanamycin 12.5-6.25 g/ml (3.125g/mL doesn't inhibit) 50 g/ml Sensitive MIC info from Barrick Lab results (G.S.)
Spectinomycin ? 60 g/ml Sensitive Spontaneous resistance
Streptomycin 6.25-12.5 g/ml (escapes observed at 6.25g/mL) 25 g/ml Sensitive Barrick Lab results (G.S.)
Chloramphenicol 4.0 g/ml 20 g/ml Sensitive Spontaneous resistance
Gentamycin 1.3 g/ml 5 g/ml Sensitive  
Tetracycline 1.0 g/ml 10 g/ml Sensitive  
Rifampicin 0.6 g/ml 100 g/ml Sensitive Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks) 1.9 g/ml (no growth at 24 hr, but growth at 48 hr) (1 g/ml doesn't inhibit) 3.75 g/ml Sensitive Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06


Common high-copy number E. coli plasmids do not work in ADP1. A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Origin Copy Number Example
ColE1 (pUC, pBR322, etc.) type plasmids Do not replicate NA
pBAV1K Low-medium copy number pBAV1K-T5-gfp
RSF1010 Low-medium copy number pBTK403
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Contributors to this topic Edit topic JeffreyBarrick, IsaacGifford
Topic revision: r2 - 2022-03-18 - 12:05:01 - Main.JeffreyBarrick
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