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Elymus nutans high molecular weight glutelin subunit gene and application thereof

A glutenin subunit, high molecular weight technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., to improve the quality of wheat

Inactive Publication Date: 2011-02-16
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report of high-quality and high-weight glutenin subunits cloned from old awn wheat. Therefore, high-quality high-molecular-weight glutenin subunits are cloned and identified from old awn wheat, and then transformed by distant hybridization and genetic engineering. In common wheat, it has important practical significance for the improvement of wheat quality

Method used

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  • Elymus nutans high molecular weight glutelin subunit gene and application thereof
  • Elymus nutans high molecular weight glutelin subunit gene and application thereof
  • Elymus nutans high molecular weight glutelin subunit gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 The high-molecular-weight glutenin subunit gene of multi-leaf old mans wheat, which is the gene encoding Sty8.1, has the nucleotide sequence shown in the 1st to 1824th positions of SEQ ID NO.1 in the sequence listing.

[0025] 1. The cloning of the high molecular weight glutenin subunit Sty8.1 gene of multi-leaf old awn wheat.

[0026] 1. Primer design:

[0027] Design PCR primers according to the protein N-terminal and C-terminal conserved sequences and published gene sequences, as follows:

[0028] P15'-ATGGCTAAGCGG(C / T)T(A / G)GTCCTCTTG-3'

[0029] P25′-CTATCACTGGCT(A / G)GCCGACAATGCG-3′

[0030] 2. Genomic DNA extraction by CTAB (cetyltrimethylammonium bromide) method:

[0031] (1) Take 50 mg of fresh multi-leaf old mangosteen leaves and grind them into powder with liquid nitrogen and quartz sand.

[0032](2) Put the powder into a centrifuge tube, add 500 μl 2×CTAB extract (2% CTAB: 1.4mol / L NaCl; 0.1mol / L Tris-HCl; 0.1mol / L EDTA, pH=8.0;) and 1 μl β - Me...

Embodiment 2

[0091] Example 2 The high-molecular-weight glutenin subunit gene of multi-leaf old awn wheat, which is the gene encoding Sty8.1, has one or more bases from the 1st to 1824th positions of SEQ ID NO.1 in the sequence listing A functionally active allele resulting from a deletion, insertion, or substitution.

[0092] The cloning of the high-molecular-weight glutenin subunit Sty8.1 gene of the multi-leaf old awn wheat and the prokaryotic expression of the high-molecular-weight glutenin subunit Sty8.1 gene of the multi-leaf old awn wheat are the same as in Example 1.

Embodiment 3

[0093] Example 3 The high-molecular-weight glutenin subunit gene of multi-leaf old awn wheat, which is a gene encoding Sty8.1, has a protein having the amino acid residue sequence shown in the first to 606th positions of SEQ ID NO.2 in the sequence listing .

[0094] The cloning of the high-molecular-weight glutenin subunit Sty8.1 gene of the multi-leaf old awn wheat and the prokaryotic expression of the high-molecular-weight glutenin subunit Sty8.1 gene of the multi-leaf old awn wheat are the same as in Example 1.

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Abstract

The invention relates to an Elymus nutans high molecular weight glutelin subunit gene and an application thereof. The gene in the invention is a Sty 8.1 coding gene with a nucleotide sequence as shown in SEQ ID NO.1 or has an allelic gene which is provided with functional activity as shown in SEQ ID NO.1 in a sequence table and is formed by deletion, insertion or substitution of one or more basicgroups, or has a protein with an amino acid residue sequence as shown in SEQ ID NO.2, or has a protein which is provided with an amino acid residue sequence as shown in SEQ ID NO.2 in a sequence table, is formed by substitution, deletion or addition of one or more amino acid residues, has the same activity as the amino acid residue sequence shown in SEQ ID NO.2, and is derived from SEQ ID NO.2. The gene obtained by the invention has a secondary structure superior to that of a high-quality gene 1Dy10, and can play an important role in improvement of wheat quality.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, in particular to a high-molecular-weight glutenin subunit gene of multi-leaf old awn wheat and the application of the gene to improve wheat quality through biotechnological methods. Background technique [0002] Wheat endosperm storage proteins can be divided into gliadins and glutenins, both of which make wheat dough extensible and elastic, and thus can be processed into various foods, among which glutenin determines the elasticity of dough Can be divided into high molecular weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW-GS) (Payne et al., 1985; Gupta and Shepherd, 1990). Although high molecular weight glutenin subunits account for only a small fraction (approximately 12%) of the total protein in wheat seeds (Halford et al., 1992), they contribute up to 70% to bread baking quality (Payne, 1987) . In common hexaploid wheat, the high molecular weig...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29A01H5/00
Inventor 王海庆刘永安窦全文陈志国
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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