Corn protein relevant to genus potyvirus virus infection and encoding gene and application thereof

A technology that encodes genes and potatoes, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as chlorosis symptoms

Inactive Publication Date: 2011-09-14
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Constitutive expression of the VI gene of cauliflower mosaic virus (CaMV) can induce severe chlorosis symptoms in transgenic plants, which are very similar to the symptoms after infection by cauliflower mosaic virus. Through differential analysis, it was found that the analogue of the transgenic VI gene Several genes in Arabidopsis thaliana and cauliflower mosaic virus infected Arabidopsi

Method used

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  • Corn protein relevant to genus potyvirus virus infection and encoding gene and application thereof
  • Corn protein relevant to genus potyvirus virus infection and encoding gene and application thereof
  • Corn protein relevant to genus potyvirus virus infection and encoding gene and application thereof

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Experimental program
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Effect test

Embodiment 1

[0036] The acquisition of embodiment 1, ZmTrm2

[0037] 1. Construction of maize suppression subtractive hybridization library

[0038] 1. Extraction of corn total RNA

[0039] 1) Take 0.1g of fresh corn leaves, grind them into powder with liquid nitrogen, transfer to a sterilized 1.5ml centrifuge tube, then add 1ml of Tri-Reagent, shake vigorously or add 1ml of Tri-Reagent to In 1×106 protoplasts frozen at -80°C, shake vigorously for 3 minutes; place on ice for 5 minutes.

[0040] 2) The homogenate was centrifuged at 4°C and 12,000 g for 10 min in an IEC desktop refrigerated centrifuge to remove insoluble components, and the supernatant was transferred to a new 1.5 ml centrifuge tube.

[0041] 3) Let stand at room temperature for 5 minutes, add 0.2ml of chloroform, shake vigorously for 15 sec, then stand at room temperature for 2-5 minutes, and then centrifuge at 4°C and 12,000×g for 15 minutes.

[0042] 4) Transfer the upper aqueous phase to a new 1.5ml centrifuge tube, a...

Embodiment 2

[0183] Example 2, ZmTrm2 protein characterization and its application in virus prevention and treatment

[0184] 1. Localization of ZmTrm2 in maize mesophyll cells

[0185] 1. Construction of ZmTrm2 expression vector

[0186] Using the sequence 2 in the artificially synthesized sequence list as a template, primers were designed to amplify the full-length reading frame of ZmTrm2, the PCR product was digested by SalI / SacI double enzymes and a 516bp gene fragment was recovered, and the pcEmGFP-DEST vector was simultaneously digested with SalI / SacI double enzymes (purchased from Invitrogen, the product catalog number is V35520), recover the large vector fragment, and connect the recovered vector large fragment with the recovered 516bp gene fragment to obtain the target plasmid. The target plasmid was transformed into E. coli, resistance screening, positive clones were picked, and the positive clones were cultured in liquid, and the positive clone plasmids were extracted for seque...

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Abstract

The invention discloses a corn protein relevant to genus potyvirus virus infection and an encoding gene and application thereof. The protein relevant to genus potyvirus virus infection, provided by the invention, is a protein shown as (a) or (b), wherein (a) is constituted by an amino acid sequence shown as a sequence 1 in a sequence table, and (b) is obtained by performing substitution and/or deletion and/or addition of one or more amino acid residues on an amino acid residue sequence shown as a sequence 1 in the sequence table, is relevant to genus potyvirus virus infection and is derived from (a). As proved by a semi-quantitative RT-PCR (Reverse Transcriptase Polymerase Chain Reaction), system infection of a sugarcane mosaic virus is facilitated by instantaneous silence of ZmTrm2 on a corn leaf, and the replication of the tobacco vein bandingmosaic viruses can be suppressed by expressing the ZmTrm2 on ordinary tobacco.

Description

technical field [0001] The present invention relates to corn protein related to potato Y virus infection, its encoding gene and application. Background technique [0002] Intermolecular interactions between plant viruses and hosts have made great progress in the past decade. However, how virus infection affects plant cells, tissues and even whole plants at the biochemical and physiological levels is still a gap in the study of plant virology. Much remains to be learned about how the virus uses its few proteins to infect plants and cause symptoms. Symptoms caused by viruses in plants are caused by physiological and structural changes in plants at the cellular level combined with slowing of plant growth and development. Through the application of recombinant DNA technology to study the role of virus gene products in the process of symptom generation, we gradually understand the process of virus infection in plants. The current research methods mainly include the following: ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/11A01N65/44A01P1/00C12N15/82A01H5/00C12R1/91C12R1/19
Inventor 范在丰施艳曹言勇周涛
Owner CHINA AGRI UNIV
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