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Application of RING1 protein for improving RDV (rice dwarf virus) resistance of plants

A dwarf disease and protein technology, applied in the application field of RING1 protein in improving plant resistance to rice dwarf virus, can solve problems such as unmentioned interaction

Inactive Publication Date: 2013-07-10
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the ubiquitination degradation of proteins is specific, and its specificity is determined by the E3 enzymes that interact with the protein. Compared with the large number of E3 enzymes that exist in organisms, the correspondence between the currently known E3 enzymes and their substrates The relationship is only lightly reported
Previous studies on RDV have never mentioned the virus's interaction with any rice E3 enzymes, nor have any reports indicated that any rice E3 enzymes have anti-RDV effects

Method used

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  • Application of RING1 protein for improving RDV (rice dwarf virus) resistance of plants
  • Application of RING1 protein for improving RDV (rice dwarf virus) resistance of plants
  • Application of RING1 protein for improving RDV (rice dwarf virus) resistance of plants

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Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1, the acquisition of RING1 protein and its coding gene

[0047] 1. Acquisition of RING1 protein and its coding gene

[0048] The kit used for the construction of the rice library was HybriZAP-2.1Two-Hybrid Predigested Vector / Gigapack Cloning Kit (Stratagene), and the library construction and screening methods were carried out according to the instructions. The bait clone pGBK-S2 used is the same as the one published in Patent No. ZL200510114386.X. The yeast strain used was AH109. The screened positive clones were sequenced and sequence analyzed, and it was determined that the encoded protein fragment had 12-308 amino acids of the amino acid sequence shown in Sequence 1 in the Sequence Listing, and the full length of the protein (ie, the protein corresponding to Sequence 1) was named RING1 , its coding gene is the nucleotide shown in Sequence 2 in the sequence listing, and it is named RING1.

[0049] 2. The cloning of the full-length sequence of RING1 and ...

Embodiment 2

[0054] Example 2. Demonstrating the interaction between RING1 protein and RDV P2 in vitro

[0055] 1. Yeast two-hybrid experiment proves the interaction between RING1 and RDV P2

[0056] The recombinant vectors pGAD-RING1 and pGBK-S2 obtained in Example 1 (S2 is the coding gene of RDV P2 protein, and the coding gene of RDV P2 protein is introduced into pGBK to obtain. The non-patent literature describing the vector is Zhu et al., 2005. The Rice Dwarf Virus P2Protein interacts with ent-Kaurene Oxidases in vivo, leading to reduced biosynthesis of Gibberellins and rice dwarf symptoms. Plant Physiology. 139:1935-1945; the public can obtain from Peking University) co-transformed yeast strain AH109 (purchase (from Clontech Company, catalog number 630444), the yeast transformation method adopts the well-known PEG / LiAc method; the transformed yeast cells are coated with auxotrophic plate SD / -Trp-Leu, and cultured at 30°C for 2 days. The transformants were streaked onto the SD / -Trp-Le...

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Abstract

The invention discloses application of RING1 protein for improving RDV (rice dwarf virus resistance) of plants, and particularly provides application of protein RING1 or a coding gene of the protein RING1 or a recombinant vector containing the coding gene for regulating RDV resistance of plants. The amino acid sequence of the protein RING1 is the sequence one in the sequence table. The experiment disclosed by the invention proves that the RING1 is E3 protein-ubiquitin ligase and capable of interacting with RDV coat protein P2 to promote the P2 to be degraded by 26S proteosome, and the cumulant of the P2 can be reduced by excessively expressing the RING1 protein in rice, thereby inhibiting the amplification of RDV and improving the anti-virus capability of rice.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of RING1 protein in improving plant resistance to rice dwarf virus. Background technique [0002] Rice dwarf virus (RDV) is a member of the genus Phytoreovirus in the Reoviridae family. RDV is the pathogen that causes rice dwarf disease, which can infect rice and cause severe rice yield reduction. The spread of RDV requires the aid of the insect vector leafhopper. RDV is a double-stranded RNA virus whose genome consists of twelve double-stranded RNAs. According to the mobility of these twelve double-stranded RNAs in polyacrylamide gel electrophoresis from slow to fast, they are named S1 to S12 respectively. The RDV genome encodes at least seven structural proteins, including P1, P2, P3, P5, P7, P8, and P9; five nonstructural proteins, including Pns4, Pns6, Pns10, Pns11, and Pns12. [0003] P2 is the coat protein of RDV, encoded by S2, which can interact with rice ...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N15/63A01H5/00
Inventor 李毅刘利芳
Owner PEKING UNIV
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