{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T07:06:24Z","timestamp":1790665584511,"version":"4.1.0"},"reference-count":52,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2019,7,1]],"date-time":"2019-07-01T00:00:00Z","timestamp":1561939200000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon 2020 research and innovation programme","award":["634942"],"award-info":[{"award-number":["634942"]}]},{"name":"European Research Council","award":["670216"],"award-info":[{"award-number":["670216"]}]},{"name":"MYCOCHASSIS"},{"name":"Spanish Ministry of Economy, Industry and Competitiveness"},{"name":"Centro de Excelencia Severo Ochoa"},{"name":"CERCA Programme\/Generalitat de Catalunya"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Mycoplasmas are important model organisms for Systems and Synthetic Biology, and are pathogenic to a wide variety of species. Despite their relevance, many of the tools established for genome editing in other microorganisms are not available for Mycoplasmas. The Tn4001 transposon is the reference tool to work with these bacteria, but the transformation efficiencies (TEs) reported for the different species vary substantially. Here, we explore the mechanisms underlying these differences in four Mycoplasma species, Mycoplasma agalactiae, Mycoplasma feriruminatoris, Mycoplasma gallisepticum and Mycoplasma pneumoniae, selected for being representative members of each cluster of the Mycoplasma genus. We found that regulatory regions (RRs) driving the expression of the transposase and the antibiotic resistance marker have a major impact on the TEs. We then designed a synthetic RR termed SynMyco RR to control the expression of the key transposon vector elements. Using this synthetic RR, we were able to increase the TE for M. gallisepticum, M. feriruminatoris and M. agalactiae by 30-, 980- and 1036-fold, respectively. Finally, to illustrate the potential of this new transposon, we performed the first essentiality study in M. agalactiae, basing our study on more than 199,000 genome insertions.<\/jats:p>","DOI":"10.1093\/dnares\/dsz012","type":"journal-article","created":{"date-parts":[[2019,5,16]],"date-time":"2019-05-16T19:16:44Z","timestamp":1558034204000},"page":"327-339","source":"Crossref","is-referenced-by-count":25,"title":["SynMyco transposon: engineering transposon vectors for efficient transformation of minimal genomes"],"prefix":"10.1093","volume":"26","author":[{"given":"Ariadna","family":"Montero-Blay","sequence":"first","affiliation":[{"name":"Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel","family":"Miravet-Verde","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Lluch-Senar","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5263-3702","authenticated-orcid":false,"given":"Carlos","family":"Pi\u00f1ero-Lambea","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Serrano","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona, Spain"},{"name":"Universitat Pompeu Fabra (UPF), Barcelona, Spain"},{"name":"ICREA, Pg. Lluis Companys 23, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2019,6,30]]},"reference":[{"key":"2019091012295654400_dsz012-B1","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1073\/pnas.77.1.494","article-title":"Phylogenetic analysis of the mycoplasmas","volume":"77","author":"Woese","year":"1980","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2019091012295654400_dsz012-B2","doi-asserted-by":"crossref","first-page":"4420","DOI":"10.1093\/nar\/24.22.4420","article-title":"Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae","volume":"24","author":"Himmelreich","year":"1996","journal-title":"Nucleic Acids Res"},{"key":"2019091012295654400_dsz012-B3","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1126\/science.1176951","article-title":"Transcriptome complexity in a genome-reduced bacterium","volume":"326","author":"Guell","year":"2009","journal-title":"Science"},{"key":"2019091012295654400_dsz012-B4","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1038\/msb.2011.38","article-title":"Quantification of mRNA and protein and integration with protein turnover in a bacterium","volume":"7","author":"Maier","year":"2011","journal-title":"Mol. Syst. Biol"},{"key":"2019091012295654400_dsz012-B5","doi-asserted-by":"crossref","first-page":"e1003191","DOI":"10.1371\/journal.pgen.1003191","article-title":"Comprehensive methylome characterization of Mycoplasma genitalium and Mycoplasma pneumoniae at single-base resolution","volume":"9","author":"Lluch-Senar","year":"2013","journal-title":"PLoS Genet"},{"key":"2019091012295654400_dsz012-B6","doi-asserted-by":"crossref","first-page":"780","DOI":"10.15252\/msb.20145558","article-title":"Defining a minimal cell: essentiality of small ORFs and ncRNAs in a genome-reduced bacterium","volume":"11","author":"Lluch-Senar","year":"2015","journal-title":"Mol. Syst. Biol"},{"key":"2019091012295654400_dsz012-B7","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.cell.2012.05.044","article-title":"A whole-cell computational model predicts phenotype from genotype","volume":"150","author":"Karr","year":"2012","journal-title":"Cell"},{"key":"2019091012295654400_dsz012-B8","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1073\/pnas.0510013103","article-title":"Essential genes of a minimal bacterium","volume":"103","author":"Glass","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2019091012295654400_dsz012-B9","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/j.1365-2958.2008.06262.x","article-title":"Large-scale transposon mutagenesis of Mycoplasma pulmonis","volume":"69","author":"French","year":"2008","journal-title":"Mol. 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A PP2C-type phosphatase serves to dephosphorylate HPr (Ser-P)","volume":"281","author":"Halbedel","year":"2006","journal-title":"J. Biol. Chem"},{"key":"2019091012295654400_dsz012-B17","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.ijmm.2006.11.001","article-title":"Tools for the genetic analysis of Mycoplasma","volume":"297","author":"Halbedel","year":"2007","journal-title":"Int. J. Med. Microbiol"},{"key":"2019091012295654400_dsz012-B18","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1007\/BF00382099","article-title":"Tn4001: a gentamicin and kanamycin resistance transposon in Staphylococcus aureus","volume":"193","author":"Lyon","year":"1984","journal-title":"Mol. Gen. 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