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Solanum lycopersicoides dun. S1Ve1 receptor protein encoding sequence and use thereof

A tomato, anti-big technology, applied in the direction of application, use of vectors to introduce foreign genetic material, plant peptides, etc.

Inactive Publication Date: 2004-12-29
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Before the present invention was published, there was no disclosure or report of the tomato-like Solanum anti-Verticillium dahliae receptor protein SlVe1 sequence and its nucleic acid sequence mentioned in this patent application

Method used

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  • Solanum lycopersicoides dun. S1Ve1 receptor protein encoding sequence and use thereof
  • Solanum lycopersicoides dun. S1Ve1 receptor protein encoding sequence and use thereof
  • Solanum lycopersicoides dun. S1Ve1 receptor protein encoding sequence and use thereof

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Embodiment 1

[0040] Cloning of the Receptor Protein (SlVe1) Gene of Solanum Solanum Resistant to Verticillium dahliae

[0041] 1. Plant material and RNA isolation (Plant material and RNA isolation)

[0042] The seeds of the tomato-like solanaceous line LA2386 were kindly donated by Dr. Chetelat of the C.M.Rick Tomato Genetic Resources Center (C.M.Rick Tomato Genetic Resources Center, Dept. Vegetable Crops, University of California, Davis, USA), Department of Vegetable Crops, University of California, Davis, USA. Seeds were soaked in 15% sodium hypochlorite disinfectant for 30 minutes, rinsed in sterilized distilled water with 4 changes of water for 4 hours, and then inoculated in the middle of several layers of sterilized filter paper at 25°C to germinate, and the germinated seeds were sown to a concentration of 1:1:1 In the culture tanks of loam, vermiculite and perlite, keep them at 20°C and 14h light / 10h dark conditions until they emerge and grow, and inoculate the regenerated plants wi...

Embodiment 2

[0057] Sequence Information and Homology Analysis of the Receptor Protein (SlVe1) Gene of Solanum Solanum Resistant to Verticillium dahliae

[0058] The novel tomato-like solanum anti-Verticillium dahliae receptor protein (SlVe1) full-length cDNA has a length of 3400bp (excluding polyA), and the detailed sequence is shown in SEQ ID NO.3, wherein the open reading frame is located at the 23-3175 core glycosides. The amino acid sequence encoded by SlVe1 was deduced according to the full-length cDNA, with a total of 1050 amino acid residues, a molecular weight of 116.97kDa, and an isoelectric point (pI) of 5.22. See SEQ ID NO.3 for the detailed sequence.

[0059] The full-length cDNA sequence of tomato-like Solanum anti-Verticillium dahliae receptor protein and its encoded protein were published in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+ Nucleotide and protein homology searches were carried out in the PIR database, ...

Embodiment 3

[0061] Copy Number Analysis of Solanum Solanum Anti-Verticillium dahliae Receptor Protein (SlVe1) Gene in Solanum Solanum

[0062] Extract DNA from tomato-like Solanaceae leaves by conventional methods (refer to "Molecular Cloning", Sambrook et al., 1989), digest the DNA with DraI, EcoRV and HindIII respectively [13 μg (microgram) / sample], transfer the DNA to the hybridization membrane (nylon film). Using Gene Images from Amersham Pharmacia TM Contents CDP-Star TMThe labeling module (PRN3540) hybridization and detection system used the partial coding sequence of the SlVe1 gene (685 bp, obtained by PCR amplification of tomato-like Solanum cDNA using FVE685 and RVE685 as primers) as a probe to hybridize the hybridization membrane (hybridize at 60°C for 16 hours). The membrane was taken out, placed in washing solution I (1*SSC, 1% SDS), and rinsed three times at 60° C. for 15 minutes each time. Transfer to washing solution II (0.1*SSC, 1% SDS) and rinse at 60°C for 3 times, e...

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Abstract

The present invention provides one new kind dahlia Streptoverticillium resisting receptor protein (S1Vc1) expressed in Solanum lycopersicoides Dun. and its coding sequence and application in improving plant disease resistance, especially in resisting verticillium wilt. The present invention relates to fusion gene constituent and new recombinant expression vector carrying the constituent, as well as transgenic plant generated via transforming plant cell with the expression vector and its subsequent generation, including plant seed and plant tissue. The present invention expresses the said gene in plant to obtain transgenic plant with strengthened resistance to plant diseases, especially verticillium wilt.

Description

technical field [0001] The invention relates to the fields of molecular biology, enzymology, physiology, genetic engineering and the like. Specifically, the present invention relates to a kind of anti-Verticillium dahliae Kleb. resistance protein (Ve) from Solanum lycopersicoides Dun., SlVe1] and its nucleic acid expressed in tomato tomato (Solanum lycopersicoides Dun.) sequence. The present invention also relates to the preparation method and use of the protein and nucleic acid sequence. Background technique [0002] Verticillium dahliae Kleb. caused by Verticillium dahliae Kleb. is a worldwide intractable vascular disease, which seriously damages the growth of important crops such as cotton, tomato, eggplant, potato and alfalfa. As a soil-borne disease, verticillium wilt has a wide range of hosts and serious damage. Chemical control costs are high, the effect is poor, and there are food safety and ecological security problems. Breeding Verticillium wilt-resistant crop v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/79C12N15/82
Inventor 唐克轩柴有荣孙小芬左开井李柱刚
Owner FUDAN UNIV
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