Construction of inducible genes-of-interest was accomplished by placing the respective gene under the control of the arabinose-inducible promoter PBAD inside the miniTn7 transposon TnAraC (23 (link), 63 (link)). The resulting transposons were transferred to the respective
Constructing A. baumannii Mutants and Variants
Construction of inducible genes-of-interest was accomplished by placing the respective gene under the control of the arabinose-inducible promoter PBAD inside the miniTn7 transposon TnAraC (23 (link), 63 (link)). The resulting transposons were transferred to the respective
Corresponding Organization : École Polytechnique Fédérale de Lausanne
Protocol cited in 2 other protocols
Variable analysis
- Deletion constructs of the respective genes
- Site-directed allele exchanges of pilA
- Placing the respective gene under the control of the arabinose-inducible promoter P_BAD inside the miniTn7 transposon TnAraC
- Phenotypes of the A. baumannii mutants or variants
- Use of counterselectable suicide vectors pGP704-Sac28 or pGP704Sac-kan
- Use of the aph cassette (Kan^r) as the selection marker
- Use of overlap extension PCR or Golden Gate assembly to construct the genetic modifications
- Screening for correct cloning products by colony PCR and Sanger sequencing
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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