{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T20:40:24Z","timestamp":1791319224388,"version":"4.3.1"},"reference-count":55,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2018,2,2]],"date-time":"2018-02-02T00:00:00Z","timestamp":1517529600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010100","name":"Jiho\u010desk\u00e1 Univerzita v \u010cesk\u00fdch Bud\u011bjovic\u00edch","doi-asserted-by":"publisher","award":["050\/2016\/P"],"award-info":[{"award-number":["050\/2016\/P"]}],"id":[{"id":"10.13039\/100010100","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000936","name":"Gordon and Betty Moore Foundation","doi-asserted-by":"publisher","award":["GBMF4983.01"],"award-info":[{"award-number":["GBMF4983.01"]}],"id":[{"id":"10.13039\/100000936","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Environmental Microbiology"],"published-print":{"date-parts":[[2018,3]]},"abstract":"<jats:title>Summary<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Diplonema papillatum<\/jats:italic>\n                    is the type species of diplonemids, which are among the most abundant and diverse heterotrophic microeukaryotes in the world's oceans. Diplonemids are also known for a unique form of post\u2010transcriptional processing in mitochondria. However, the lack of reverse genetics methodologies in these protists has hampered elucidation of their cellular and molecular biology. Here we report a protocol for\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    transformation. We have identified several antibiotics to which\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    is sensitive and thus are suitable selectable markers, and focus in particular on puromycin. Constructs were designed encoding antibiotic resistance markers, fluorescent tags, and additional genomic sequences from\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    to facilitate vector integration into chromosomes. We established conditions for effective electroporation, and demonstrate that electroporated constructs can be stably integrated in the\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    nuclear genome. In\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    transformants, the heterologous puromycin resistance gene is transcribed into mRNA and translated into protein, as determined by Southern hybridization, reverse transcription, and Western blot analyses. This is the first documented case of transformation in a euglenozoan protist outside the well\u2010studied kinetoplastids, making\n                    <jats:italic>D. papillatum<\/jats:italic>\n                    a genetically tractable organism and potentially a model system for marine microeukaryotes.\n                  <\/jats:p>","DOI":"10.1111\/1462-2920.14041","type":"journal-article","created":{"date-parts":[[2018,1,10]],"date-time":"2018-01-10T10:04:17Z","timestamp":1515578657000},"page":"1030-1040","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Transformation of\n                    <i>Diplonema papillatum<\/i>\n                    , the type species of the highly diverse and abundant marine microeukaryotes Diplonemida (Euglenozoa)"],"prefix":"10.1111","volume":"20","author":[{"given":"Binnypreet","family":"Kaur","sequence":"first","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"},{"name":"Faculty of Sciences University of South Bohemia \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matus","family":"Valach","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Robert\u2010Cedergren Centre for Bioinformatics and Genomics Universit\u00e9 de Montr\u00e9al Montreal Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priscila","family":"Pe\u00f1a\u2010Diaz","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandrine","family":"Moreira","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Robert\u2010Cedergren Centre for Bioinformatics and Genomics Universit\u00e9 de Montr\u00e9al Montreal Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick J.","family":"Keeling","sequence":"additional","affiliation":[{"name":"Botany Department University of British Columbia Vancouver Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gertraud","family":"Burger","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Robert\u2010Cedergren Centre for Bioinformatics and Genomics Universit\u00e9 de Montr\u00e9al Montreal Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julius","family":"Luke\u0161","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"},{"name":"Faculty of Sciences University of South Bohemia \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Drahom\u00edra","family":"Faktorov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"},{"name":"Faculty of Sciences University of South Bohemia \u010cesk\u00e9 Bud\u011bjovice (Budweis) Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,2,2]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.2012.00644.x"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0163049"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkm249"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1098\/rsos.170095"},{"key":"e_1_2_7_6_1","first-page":"333","article-title":"Transfection of Leishmania and Trypanosoma brucei by electroporation","volume":"21","author":"Beverley S.M.","year":"1993","journal-title":"Methods Mol Biol"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-10-145"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1261605"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep22252"},{"key":"e_1_2_7_10_1","doi-asserted-by":"crossref","unstructured":"Faktorov\u00e1 D. Valach M. Kaur B. Burger G. andLuke\u0161 J.(2018) Mitochondrial RNA editing and processing in diplonemid protists. \u201cRNA Metabolism in Mitochondria\u201d Springer series \u201cNucleic Acids and Molecular Biology\u201d in press.","DOI":"10.1007\/978-3-319-78190-7_6"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2007.09.007"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2016.09.031"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2016.09.013"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10295-011-1032-6"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.13830"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1360-1385(99)01430-2"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00508-13"},{"key":"e_1_2_7_18_1","first-page":"1","article-title":"Ueber marine Flagellaten","volume":"32","author":"Griessman K.","year":"1914","journal-title":"Arch Protistenkd"},{"key":"e_1_2_7_19_1","first-page":"117","article-title":"Optimizing electroporation conditions for the transformation of mammalian cells","volume":"130","author":"Heiser W.C.","year":"2000","journal-title":"Methods Mol Biol"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1142"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/wrna.1124"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.4161\/rna.23340"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0711799105"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00386-09"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/jeu.12338"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2008.01737.x"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1080\/00222939000770571"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2015.04.052"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.protis.2010.11.003"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1148033"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1128\/EC.4.6.1137-1146.2005"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.052308.093131"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2016.0520"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw188"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.mi.25.100171.002415"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.1973.tb00895.x"},{"key":"e_1_2_7_37_1","article-title":"Transfection by electroporation","author":"Potter H.","year":"2003","journal-title":"Curr Protoc Mol Biol"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0001-706X(97)00079-X"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.2007.00244.x"},{"key":"e_1_2_7_40_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.17.1.267"},{"key":"e_1_2_7_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-9365(11)80225-3"},{"key":"e_1_2_7_42_1","first-page":"141","article-title":"Ultrastructure of a flagellate, Isonema nigricans nov. gen. nov. sp., from a polluted marine habitat","volume":"4","author":"Schuster F.L.","year":"1968","journal-title":"Protistologica"},{"key":"e_1_2_7_43_1","doi-asserted-by":"publisher","DOI":"10.1155\/2014\/893272"},{"key":"e_1_2_7_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-9365(97)80012-7"},{"key":"e_1_2_7_45_1","first-page":"5","article-title":"Taxonomie des phytoplanktons einiger seen in Uppland, Schweden","volume":"9","author":"Skuja H.","year":"1948","journal-title":"Symb Bot Upsal"},{"key":"e_1_2_7_46_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.7.2554-2559.1998"},{"key":"e_1_2_7_47_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.2001.tb00321.x"},{"key":"e_1_2_7_48_1","doi-asserted-by":"crossref","unstructured":"Tashyreva D. 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