Optimizing CaCl 2 -mediated transformation of Pseudomonas aeruginosa SDK-6 with pJN105 using OFAT: A novel and efficient cloning approach.

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  • Author(s): Kaur D;Kaur D;Kaur D; Singh V; Singh V; Gupta S; Gupta S
  • Source:
    Current genetics [Curr Genet] 2024 Jul 31; Vol. 70 (1), pp. 11. Date of Electronic Publication: 2024 Jul 31.
  • Publication Type:
    Journal Article
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Springer International Country of Publication: United States NLM ID: 8004904 Publication Model: Electronic Cited Medium: Internet ISSN: 1432-0983 (Electronic) Linking ISSN: 01728083 NLM ISO Abbreviation: Curr Genet Subsets: MEDLINE
    • Publication Information:
      Publication: New York Ny : Springer International
      Original Publication: [New York] Springer International.
    • Subject Terms:
      Pseudomonas aeruginosa*/genetics ; Cloning, Molecular* ; Transformation, Bacterial* ; Plasmids*/genetics ; Genetic Vectors*/genetics; Calcium Chloride
    • 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 × 10 5 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.
      (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
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    • Contributed Indexing:
      Keywords: Pseudomonas aeruginosa; Heat shock; OFAT; Transformants; pJN105
    • Accession Number:
      M4I0D6VV5M (Calcium Chloride)
    • Publication Date:
      Date Created: 20240731 Date Completed: 20240731 Latest Revision: 20241213
    • Publication Date:
      20241213
    • Accession Number:
      10.1007/s00294-024-01295-5
    • Accession Number:
      39083080