{
  "abstract": "Genetic manipulation of mycobacteria remains critical in functional genomics and modern microbiology. However, this process is often laborious due to limited cloning flexibility and low throughput. Here, we describe a streamlined cloning platform for expressing a gene of interest either alone or fused to a monomeric fluorescent protein under the control of two promoters of choice. This platform combines three simple PCR amplification reactions and four simultaneous ligations. Accordingly, this four-step procedure provides a fast and flexible cloning strategy for protein expression and subcellular localization analyses in mycobacteria. This cost-effective approach is compatible with some commonly used mycobacterial vectors and host strains. Using M. tuberculosis triacylglycerol synthase 1 gene (tgs1) as a proof of concept, we demonstrate the efficient cloning, robust expression, and reliable localization of the resulting fusion protein. This platform significantly reduces hands-on time and experimental complexity, providing a practical tool for functional and microbiological studies in mycobacteria.",
  "abstract_shown": true,
  "affiliations": [],
  "article_type": null,
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "name": "Maria Sofia Deschutter"
    },
    {
      "name": "Eva Beaumale"
    },
    {
      "name": "Stéphane Canaan"
    },
    {
      "name": "Gabriela Gago"
    },
    {
      "name": "Pierre Santucci"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/5w04da71f4sm5.bib",
  "cited_by": 0,
  "doi": "10.1016/j.biochi.2026.08.008",
  "fulltext_url": null,
  "id": "5w04da71f4sm5",
  "issns": [
    "0300-9084",
    "1638-6183"
  ],
  "landing_url": "https://hal.science/hal-05844486v1",
  "language": "en",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "hal",
      "0.sdv"
    ],
    [
      "openalex",
      "2725|Tuberculosis Research and Epidemiology"
    ]
  ],
  "openalex": "W7204073777",
  "pdf_url": "https://hal.science/hal-05844486/document",
  "pmcid": null,
  "pmid": null,
  "published": "2026-08-28",
  "publisher": "Elsevier",
  "ris": "https://research.draconitic.com/paper/5w04da71f4sm5.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:03Z",
      "native_id": "hal-05844486",
      "source": "hal",
      "url": "https://hal.science/hal-05844486v1"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "openalex:2725|Tuberculosis Research and Epidemiology",
      "subfield": "infectious-disease"
    }
  ],
  "title": "A simple multi-cloning strategy for gene expression and subcellular localization in mycobacteria",
  "title_text": "A simple multi-cloning strategy for gene expression and subcellular localization in mycobacteria",
  "type": "peer-reviewed",
  "updated_at": "2026-10-31T12:20:56Z",
  "url": "https://research.draconitic.com/paper/5w04da71f4sm5",
  "venue": "Biochimie",
  "versions": []
}