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Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120

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    SYSNO ASEP0537734
    Document TypeJ - Journal Article
    R&D Document TypeJournal Article
    Subsidiary JČlánek ve WOS
    TitleConstruction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120
    Author(s) Srivastava, Amit (MBU-M) ORCID
    Ballal, A. (IN)
    Forchhammer, K. (DE)
    Tripathi, A. K. (IN)
    Article numbere3528
    Source TitleBio-protocol - ISSN 2331-8325
    Roč. 10, č. 4 (2020)
    Number of pages14 s.
    Languageeng - English
    CountryUS - United States
    KeywordsAntisenseRNA (asRNA) ; Knock-down ; Anabaena PCC 7120 ; Cyanobacteria ; Green fluorescent protein ; psbA1 promoter
    Subject RIVEE - Microbiology, Virology
    OECD categoryMicrobiology
    R&D ProjectsLO1416 GA MŠk - Ministry of Education, Youth and Sports (MEYS)
    Method of publishingOpen access
    Institutional supportMBU-M - RVO:61388971
    UT WOS000583091400009
    DOI10.21769/BioProtoc.3528
    AnnotationAnabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a bottleneck in the elucidation of its genomic function. Antisense RNA (asRNA) mediated approach renders the study of essential genes possible by suppressing (but not completely eliminating) expression of the target gene, thus allowing them to function to some extent. Recently, we have successfully implemented this approach using the strong endogenous promoter of the psbAl gene (D1 subunit of Photosystem II) introduced into a highcopy replicative plasmid (pAM1956) to suppress the transcript level of the target gene alr0277 (encoding a sigma factor, SigJ/Alr0277) in Anabaena. This protocol represents an efficient and easy procedure to further explore the functional genomics, expanding the scope of basic and applied research in these ecologically important cyanobacteria.
    WorkplaceInstitute of Microbiology
    ContactEliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
    Year of Publishing2021
    Electronic addresshttps://bio-protocol.org/e3528?sid=f85bba85-5c8b-4cd0-96d2-e2c9090f9d97&sousouid=20210118012731766&ptype=0&px=1&intwzid=3528
Number of the records: 1