Name: virus-induced gene silencing (vigs) in nicotiana benthamiana and tomato
Uploaded: 2009-06-10T00:00:00
Description: Watch this Scientific Journal Video about Virus-induced Gene Silencing VIGS in Nicotiana benthamiana and Tomato at JoVE.com

हम पांडुलिपि पर उसे बहुमूल्य अंतर्दृष्टि के लिए धन्यवाद डा. पेट्रीसिया Manosalva. अनुदान राष्ट्रीय विज्ञान फाउंडेशन प्लांट जीनोम कार्यक्रम, पुरस्कार संख्या DBI 0605059 द्वारा प्रदान की गई थी.

भाग 1: संयंत्र सामग्री एन benthamiana मुंह बंद करने के लिए इस्तेमाल पौधों लगभग 2 आधा पुराने सप्ताह जो समय है जब cotyledons और पहले 2 है चाहिए - 4 सच पत्तियों में उभरा है . 8 दिनों के बाद उद्भव, जब सच पत्ते अभी तक ?...

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वायरस प्रेरित जीन मुंह बंद एक तरीका है कि तेजी से रिवर्स आनुवंशिक स्क्रीन की अनुमति देता है. यह टी डीएनए या transposon मध्यस्थता जीन दस्तक बहिष्कार, जो केवल Arabidopsis और मक्का जैसे कुछ पौधों में उपलब्ध हैं की पी?...

virus-induced gene silencing (vigs) in nicotiana benthamiana and tomato

RNA interference (RNAi) is a highly specific gene-silencing phenomenon triggered by dsRNA1. This silencing mechanism uses two major classes of RNA regulators: microRNAs, which are produced from non-protein coding genes and short interfering RNAs (siRNAs). Plants use RNAi to control transposons and to exert tight control over developmental processes such as flower organ formation and leaf development2,3,4. Plants also use RNAi to defend themselves against infection by viruses. Consequently, many viruses have evolved suppressors of gene silencing to allow their successful colonization of their host5.
Virus-induced gene silencing (VIGS) is a method that takes advantage of the plant RNAi-mediated antiviral defense mechanism. In plants infected with unmodified viruses the mechanism is specifically targeted against the viral genome. However, with virus vectors carrying sequences derived from host genes, the process can be additionally targeted against the corresponding host mRNAs. VIGS has been adapted for high-throughput functional genomics in plants by using the plant pathogen Agrobacterium tumefaciens to deliver, via its Ti plasmid, a recombinant virus carrying the entire or part of the gene sequence targeted for silencing. Systemic virus spread and the endogenous plant RNAi machinery take care of the rest. dsRNAs corresponding to the target gene are produced and then cleaved by the ribonuclease Dicer into siRNAs of 21 to 24 nucleotides in length. These siRNAs ultimately guide the RNA-induced silencing complex (RISC) to degrade the target transcript2.
Different vectors have been employed in VIGS and one of the most frequently used is based on tobacco rattle virus (TRV). TRV is a bipartite virus and, as such, two different A. tumefaciens strains are used for VIGS. One carries pTRV1, which encodes the replication and movement viral functions while the other, pTRV2, harbors the coat protein and the sequence used for VIGS6,7. Inoculation of Nicotiana benthamiana and tomato seedlings with a mixture of both strains results in gene silencing. Silencing of the endogenous phytoene desaturase (PDS) gene, which causes photobleaching, is used as a control for VIGS efficiency. It should be noted, however, that silencing in tomato is usually less efficient than in N. benthamiana. RNA transcript abundance of the gene of interest should always be measured to ensure that the target gene has efficiently been down-regulated. Nevertheless, heterologous gene sequences from N. benthamiana can be used to silence their respective orthologs in tomato and vice versa8.

Watch this Scientific Journal Video about Virus-induced Gene Silencing VIGS in Nicotiana benthamiana and Tomato at JoVE.com

Virus-induced Gene Silencing VIGS in Nicotiana benthamiana and Tomato

एक वायरस प्रेरित जीन अभिव्यक्ति की दस्तक नीचे में जीन मुंह बंद विधि (VIGS) का विवरण<em&gt; Nicotiana benthamiana</em&gt; और टमाटर.

वायरस - प्रेरित जीन (VIGS) मुंह बंद Nicotiana benthamiana टमाटर और

RNA interference (RNAi) is a highly specific gene-silencing phenomenon triggered by dsRNA1. This silencing mechanism uses two major classes of RNA ...

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Virus-induced Gene Silencing (VIGS) in Nicotiana benthamiana and Tomato

Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms

Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms

लाइव सेल कैल्शियम इमेजिंग siRNA मध्यस्थता जीन मुंह बंद के साथ संयुक्त Ca पहचानता 2 + ईआर झिल्ली में रिसाव चैनल और उनके विनियामक तंत्र

In mammalian cells, the endoplasmic reticulum (ER) plays a key role in protein biogenesis as well as in calcium signalling1. The heterotrimeric Sec61 ...

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प्रेरण और सेल संस्कृति में Hypoxia का परीक्षण

Hypoxia is defined as the reduction or lack of oxygen in organs, tissues, or cells.  This decrease of oxygen tension can be due to a reduced supply in ...

Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton

Agrobacterium की मध्यस्थता वायरस प्रेरित कपास जीन का मुंह बंद परख

Cotton (Gossypium hirsutum) is one of the most important crops worldwide. Considerable efforts have been made on molecular breeding of new varieties.  The ...

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Ribosome बाध्य नवजात polypeptides का अलगाव<em&gt; इन विट्रो में</em&gt; MRNA के साथ translational रोकें ब्लॉग की पहचान के लिए

The rate of translational elongation is non-uniform. mRNA secondary structure, codon usage and mRNA associated proteins may alter ribosome movement on the ...

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

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Extensive research has provided ample evidences suggesting that protein folding in the cell is a co-translational process1-5. However, the exact pathway ...

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

आलू और निकोटियाना बेंथामियाना में एग्रोइंट्रिकेशन और पीवीएक्स एग्रोइंफेक्शन

Agroinfiltration and PVX agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. The most ...

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

अनुकूलन और की उपयोगिता<em&gt; Agrobacterium</emमें&gt; मध्यस्थता क्षणिक प्रोटीन उत्पादन<em&gt; निकोटियाना</em

Agrobacterium-mediated transient protein production in plants is a promising approach to produce vaccine antigens and therapeutic proteins within a short ...

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

रोगवाहक मच्छरों का लार्वा में chitosan / दखल शाही सेना nanoparticle के मध्यस्थता जीन मुंह बंद

Vector mosquitoes inflict more human suffering than any other organism&#8212;and kill more than one million people each year. The mosquito genome projects ...

Meiotic Spindle Assessment in Mouse Oocytes by siRNA-mediated Silencing

SiRNA की मध्यस्थता मुंह बंद करके माउस oocytes में meiotic तकला आकलन

Errors in chromosome segregation during meiotic division in gametes can lead to aneuploidy that is subsequently transmitted to the embryo upon ...

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Luciferase निकोटियाना benthamiana में पूरक इमेजिंग परख क्षणिक प्रोटीन प्रोटीन संपर्क गतिशीलता का निर्धारण करने के लिए छोड़ देता है

Protein-protein interactions are fundamental mechanisms for relaying signal transduction in most cellular processes; therefore, identification of novel ...