Počet záznamů: 1
Transformation of Diplonema papillatum, the type species of the highly diverse and abundant marine microeukaryotes Diplonemida (Euglenozoa)
- 1.
SYSNO ASEP 0498712 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Transformation of Diplonema papillatum, the type species of the highly diverse and abundant marine microeukaryotes Diplonemida (Euglenozoa) Tvůrce(i) Kaur, Binnypreet (BC-A) ORCID
Valach, M. (CA)
Peña-Diaz, Priscila (BC-A) ORCID, RID
Moreira, S. (CA)
Keeling, P. J. (CA)
Burger, G. (CA)
Lukeš, Julius (BC-A) RID, ORCID
Faktorová, Drahomíra (BC-A) RID, ORCIDCelkový počet autorů 8 Zdroj.dok. Environmental Microbiology. - : Wiley - ISSN 1462-2912
Roč. 20, č. 3 (2018), s. 1030-1040Poč.str. 11 s. Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova TRYPANOSOMA-BRUCEI ; GENE-EXPRESSION ; PROTEINS ; DNA ; KINETOPLASTIDA ; RECOMBINATION ; TRANSFECTION ; EUKARYOTES Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Genetics and heredity (medical genetics to be 3) CEP GA15-21974S GA ČR - Grantová agentura ČR GA16-18699S GA ČR - Grantová agentura ČR LL1601 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora BC-A - RVO:60077344 UT WOS 000428393900010 EID SCOPUS 85041280592 DOI 10.1111/1462-2920.14041 Anotace Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd Diplonema papillatum 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-transcriptional 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 D. papillatum transformation. We have identified several antibiotics to which D. papillatum 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 D. papillatum to facilitate vector integration into chromosomes. We established conditions for effective electroporation, and demonstrate that electroporated constructs can be stably integrated in the D. papillatum nuclear genome. In D. papillatum 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-studied kinetoplastids, making D. papillatum a genetically tractable organism and potentially a model system for marine microeukaryotes. Pracoviště Biologické centrum (od r. 2006) Kontakt Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Rok sběru 2019
Počet záznamů: 1