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Updated: Jun 19, 2026
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Published on: July 13, 2012
Optimizing CaCl2-mediated transformation of Pseudomonas aeruginosa SDK-6 with pJN105 using OFAT: A novel and
Damanjeet Kaur1,2, Vijay Singh3, Saurabh Gupta4
1Department of Microbiology, Mata Gujri College, Fatehgarh Sahib, Punjab, 140406, India.
Abstract:
Cloning and expression of a gene in the desired host is required for optimum production in recombinant strains. The present research is the first attempt to optimize the physiological conditions for the transformation of Pseudomonas aeruginosa SDK-6 with pJN105. Different factors, such as inoculum size, incubation period, heat shock temperature, and heat shock time were optimized using one factor at a time (OFAT) followed by the selection of transformants using gentamicin resistance marker. The maximum number of transformants (2.002 ± 0.077 × 105 cfu/ µg of plasmid DNA) were reported with 0.5% (v/v) inoculum, an incubation period of 3 h, and heat shock treatment at 50 °C for 1 min. An overall 12-fold increase in transformation efficiency was observed. The presence of a 6055 bp band on agarose gel confirmed the transformation of Pseudomonas aeruginosa with the vector pJN105.
