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A simple method for construction of artificial microRNA vector in plant
KTH, School of Biotechnology (BIO).
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2014 (English)In: Biotechnology letters, ISSN 0141-5492, E-ISSN 1573-6776, Vol. 36, no 10, 2117-2123 p.Article in journal (Refereed) Published
Abstract [en]

Artificial microRNA (amiRNA) is a powerful tool for silencing genes in many plant species. Here we provide an easy method to construct amiRNA vectors that reinvents the Golden Gate cloning approach and features a novel system called top speed amiRNA construction (TAC). This speedy approach accomplishes one restriction-ligation step in only 5 min, allowing easy and high-throughput vector construction. Three primers were annealed to be a specific adaptor, then digested and ligated on our novel vector pTAC. Importantly, this method allows the recombined amiRNA constructs to maintain the precursor of osa-miR528 with exception of the desired amiRNA/amiRNA* sequences. Using this method, our results showed the expected decrease of targeted genes in Nicotiana benthamiana and Oryza sativa.

Place, publisher, year, edition, pages
2014. Vol. 36, no 10, 2117-2123 p.
Keyword [en]
Artificial microRNA, Gene silencing, Golden gate cloning, High-throughput construction, Nicotiana benthamiana, Osa-miR528
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Other Biological Topics
Identifiers
URN: urn:nbn:se:kth:diva-153841DOI: 10.1007/s10529-014-1570-xISI: 000341486600025Scopus ID: 2-s2.0-84906948235OAI: oai:DiVA.org:kth-153841DiVA: diva2:755058
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QC 20141013

Available from: 2014-10-13 Created: 2014-10-09 Last updated: 2017-12-05Bibliographically approved

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Zhong, Sheng
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