Počet záznamů: 1  

Transformation of Diplonema papillatum, the type species of the highly diverse and abundant marine microeukaryotes Diplonemida (Euglenozoa)

  1. 1.
    SYSNO ASEP0498712
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    Poddruh JČlánek ve WOS
    NázevTransformation 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, ORCID
    Celkový počet autorů8
    Zdroj.dok.Environmental Microbiology. - : Wiley - ISSN 1462-2912
    Roč. 20, č. 3 (2018), s. 1030-1040
    Poč.str.11 s.
    Forma vydáníTištěná - P
    Jazyk dok.eng - angličtina
    Země vyd.GB - Velká Británie
    Klíč. slovaTRYPANOSOMA-BRUCEI ; GENE-EXPRESSION ; PROTEINS ; DNA ; KINETOPLASTIDA ; RECOMBINATION ; TRANSFECTION ; EUKARYOTES
    Vědní obor RIVEB - Genetika a molekulární biologie
    Obor OECDGenetics and heredity (medical genetics to be 3)
    CEPGA15-21974S GA ČR - Grantová agentura ČR
    GA16-18699S GA ČR - Grantová agentura ČR
    LL1601 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
    Institucionální podporaBC-A - RVO:60077344
    UT WOS000428393900010
    EID SCOPUS85041280592
    DOI10.1111/1462-2920.14041
    AnotaceEnvironmental 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.
    PracovištěBiologické centrum (od r. 2006)
    KontaktDana Hypšová, eje@eje.cz, Tel.: 387 775 214
    Rok sběru2019
Počet záznamů: 1  

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