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Tagging Fluorescent Reporter to Epinecidin-1 Antimicrobial Peptide

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  • Sivakumar Jeyarajan

    (Microbial Biotechnology Laboratory, Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
    Transgenic Animal Model Core, Biomedical Research Core Facilities, University of Michigan, Ann Arbor, MI 48108, USA)

  • Harini Priya Ramesh

    (Microbial Biotechnology Laboratory, Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
    Equal first authorship.)

  • Atchyasri Anbarasu

    (Swamy Vivekananda Medical College Hospital and Research Institute, Tiruchengode 637205, Tamil Nadu, India)

  • Jayasudha Jayachandran

    (Microbial Biotechnology Laboratory, Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India)

  • Anbarasu Kumarasamy

    (Microbial Biotechnology Laboratory, Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India)

Abstract

In this study, we successfully cloned the fluorescent proteins eGFP and DsRed in-frame with the antimicrobial peptide epinecidin-1 (FIFHIIKGLFHAGKMIHGLV) at the N-terminal. The cloning strategy involved inserting the fluorescent reporters into the expression vector, followed by screening for positive clones through visual fluorescence detection and molecular validation. The visually identified fluorescent colonies were confirmed as positive by PCR and plasmid migration assays, indicating successful cloning. This fusion of fluorescent reporters with a short antimicrobial peptide enables real-time visualization and monitoring of the peptide’s mechanism of action on membranes and within cells, both in vivo and in vitro. The fusion of eGFP and DsRed to epinecidin-1 did not impair the expression or fluorescence of the reporter protein.

Suggested Citation

  • Sivakumar Jeyarajan & Harini Priya Ramesh & Atchyasri Anbarasu & Jayasudha Jayachandran & Anbarasu Kumarasamy, 2025. "Tagging Fluorescent Reporter to Epinecidin-1 Antimicrobial Peptide," J, MDPI, vol. 8(4), pages 1-15, November.
    • Handle: RePEc:gam:jjopen:v:8:y:2025:i:4:p:42-:d:1785916
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