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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 ASEP 0537734 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Construction 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 number e3528 Source Title Bio-protocol. - : Bio-protocol - ISSN 2331-8325
Roč. 10, č. 4 (2020)Number of pages 14 s. Language eng - English Country US - United States Keywords AntisenseRNA (asRNA) ; Knock-down ; Anabaena PCC 7120 ; Cyanobacteria ; Green fluorescent protein ; psbA1 promoter Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects LO1416 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support MBU-M - RVO:61388971 UT WOS 000583091400009 DOI 10.21769/BioProtoc.3528 Annotation Anabaena 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. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2021 Electronic address https://bio-protocol.org/e3528?sid=f85bba85-5c8b-4cd0-96d2-e2c9090f9d97&sousouid=20210118012731766&ptype=0&px=1&intwzid=3528
Number of the records: 1