ICAR Research Data Repository for Knowledge Management
One-step DNA fragment assembly for expressing intron-containing hairpin RNA in plants for gene silencing
KRISHI: Publication and Data Inventory Repository
View Archive Info| Field | Value | |
| Title |
One-step DNA fragment assembly for expressing intron-containing hairpin RNA in plants for gene silencing
Not Available |
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| Creator |
M. Manamohan, G. Sharath Chandra, R. Asokan, H. Deepa, M.N. Prakash
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| Subject |
RNA interference, Binary vector, ihpRNA, In planta dsRNA expression, Single PCR, One-step cloning
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| Description |
Not Available
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. Not Available |
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| Date |
2019-10-19T09:32:37Z
2019-10-19T09:32:37Z 2013-02-01 |
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| Type |
Article
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| Identifier |
Not Available
0003-2697 http://krishi.icar.gov.in/jspui/handle/123456789/23656 |
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| Language |
English
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| Relation |
Not Available;
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| Publisher |
Elsevier
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