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http://krishi.icar.gov.in/jspui/handle/123456789/23656
Title: | One-step DNA fragment assembly for expressing intron-containing hairpin RNA in plants for gene silencing |
Other Titles: | Not Available |
Authors: | M. Manamohan, G. Sharath Chandra, R. Asokan, H. Deepa, M.N. Prakash |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::Indian Institute of Horticultural Research |
Published/ Complete Date: | 2013-02-01 |
Project Code: | Not Available |
Keywords: | RNA interference, Binary vector, ihpRNA, In planta dsRNA expression, Single PCR, One-step cloning |
Publisher: | Elsevier |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | Double-stranded RNA-mediated RNA interference in plants involves generating a construct expressing intron-containing hairpin RNA (ihpRNA), which usually is a cumbersome, multistep process. Here, we describe a simplified method involving single steps of PCR, restriction, ligation, and transformation for assembling an ihpRNA construct for plant transformation. Our method has several advantages over the currently available ones, viz., wider choice of restriction sites and facility for rapid screening of positive clones, among others. We demonstrate the utility of this approach in assembling the tomato phytoene desaturase gene. This simplified DNA fragment assembly strategy for ihpRNA construction facilitates high-throughput gene silencing in plants. |
Description: | Not Available |
ISSN: | 0003-2697 |
Type(s) of content: | Article |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | Analytical Biochemistry |
NAAS Rating: | 8.88 |
Volume No.: | 433(2) |
Page Number: | 189-191 |
Name of the Division/Regional Station: | Biotechnology |
Source, DOI or any other URL: | https://doi.org/10.1016/j.ab.2012.09.034 |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/23656 |
Appears in Collections: | HS-IIHR-Publication |
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