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The lethal properties of hok protein have been utilized for the environmental containment of microbes and the development of potential vaccine candidates. This study aimed to demonstrate the potent anti\u2010microbial property of a 19 amino acid (AA) long N\u2010terminal fragment of hok peptide. This was accomplished by designing a conditional suicide system based on <jats:italic>hok<\/jats:italic> gene expression cloned in an anhydrotetracycline (aTc) inducible vector \u2013 pASK75. Heat shock and electroporation were utilized for the transformation of <jats:italic>Escherichia coli<\/jats:italic> and <jats:italic>Vibrio cholerae<\/jats:italic> cells, respectively. The minimal induction concentration (MI<jats:sub>d<\/jats:sub>C) of aTc, determined by analyzing the expression of green fluorescent protein cloned separately into pASK75 vector, was 30\u2009ng\/mL. As <jats:italic>hok<\/jats:italic> gene was synthesized de novo (using recombinant polymerase chain reaction) in our study, various random sized hok fragments were generated (as a result of the error\u2010prone nature of Taq polymerase). The smallest hok fragment able to bring about effective antimicrobial killing was a 19 AA long N\u2010terminal fragment of hok having the wild type sequence, except for the carboxy terminus AA residue. The MI<jats:sub>d<\/jats:sub>C of aTc in our experiments was 6\u2010fold lower than previously reported, making our bacterial clones suitable for use in mammalian systems as potential vaccine candidates. Based on our experiments, we hypothesize the 19 AA long N\u2010terminal fragment of hok peptide to be the smallest possible hok fragment sufficient to bring about effective antimicrobial killing.<\/jats:p>","DOI":"10.1111\/1348-0421.12849","type":"journal-article","created":{"date-parts":[[2020,9,18]],"date-time":"2020-09-18T07:20:34Z","timestamp":1600413634000},"page":"737-746","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Cloning of <i>hok<\/i> gene into anhydrotetracycline inducible pASK75 vector reveals potent antimicrobial effect of 19 amino acid long N\u2010terminal fragment of hok peptide"],"prefix":"10.1111","volume":"64","author":[{"given":"Anit","family":"Kaur","sequence":"first","affiliation":[{"name":"Department of Biochemistry Post Graduate Institute of Medical Education and Research (PGIMER)  Chandigarh India"},{"name":"Allergy Immunology Unit, Department of Pediatrics, Advanced Pediatrics Centre 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