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Dehydrin Gene of Lentil Bean and Its Application

A technology of dehydrating protein and flat clover beans, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of no stress resistance function identification, etc., and achieve the effect of improving salt resistance and heat resistance

Inactive Publication Date: 2019-04-09
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 are only sporadic reports on the cloning of the dehydrin gene of fennel bean, but there is no report on the identification of its stress resistance function

Method used

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  • Dehydrin Gene of Lentil Bean and Its Application
  • Dehydrin Gene of Lentil Bean and Its Application
  • Dehydrin Gene of Lentil Bean and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 linseed bean dehydrin gene, the gene is MdHN3 The coding gene has the nucleotide sequence shown in the 1st to 890th positions of SEQID NO.1 in the sequence listing.

[0035] 1. Alfalfa bean dehydrated protein MdHN3 Gene cloning.

[0036] 1. Low temperature stress treatment and transcriptome sequencing of lentil seedlings:

[0037] ⑴ Low temperature stress treatment of lentil beans:

[0038] After treating the seeds of clover beans with concentrated sulfuric acid for 10 minutes, rinse them with distilled water several times to remove residual sulfuric acid. The above-mentioned treated seeds were placed in a double-layer filter paper petri dish, and germinated under the conditions of 21°C and 16h / 8h photoperiod; 3 days later, the germinated seedlings were transplanted in vermiculite: a mixture of nutrient soil (3:1) Continue to cultivate in the substrate under the above conditions, and irrigate once a week with nutrient solution containing 1 / 2 MS; after...

Embodiment 2

[0120] Embodiment 2 linseed bean dehydrin gene, the gene is MdHN3 The coding gene has a functionally active allele formed by the deletion, insertion or substitution of one or more bases from the first to the 890th position of SEQID NO.1 in the sequence listing.

[0121] Lentil Dehydrated Protein MdHN3 Cloning of the gene and the dehydrin of linseed bean MdHN3 The prokaryotic expression of the gene is the same as in Example 1.

Embodiment 3

[0122] Embodiment 3 Alfalfa bean dehydrin gene, the gene is MdHN3 coding gene, which has figure 2 A protein having the amino acid residue sequence shown in the first to 221st positions of SEQ ID NO.2 in the sequence listing.

[0123] Lentil Dehydrated Protein MdHN3 Cloning of the gene and the dehydrin of linseed bean MdHN3 The prokaryotic expression of the gene is the same as in Example 1.

[0124] right MdHN3 The protein product encoded by the cDNA reading frame consists of 221 amino acid residues with a molecular weight ( mw ) is 24.84 kD, the theoretical isoelectric point ( pI ) is 5.56, and it is an acidic protein. The total average hydrophobicity index Grand average of hydropathicity (GRAVY) is -1.444, which has high hydrophilicity.

[0125] The Pfam database (http: / / pfam.janelia.org) database searched the structural domain of the amino acid sequence encoded by MrDHN3, indicating that the MrDHN3 protein belongs to the LEAII protein family, and there is an S-f...

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Abstract

The invention relates to a dehydrated protein gene for medicago ruthenica (L.). The gene is an MrDHN3 encoding gene and comes from the medicago ruthenica (L.). The gene has a nucleotide sequence shown as SEQ ID NO. 1 in a sequence table, or has an allele with functional activity, or has protein of an amino acid residue sequence shown as SEQ ID NO. 2 in the sequence table, or has protein which comprises an S-fragment including a series of serine (Ser) residues and two K-fragments (EKKGIMDKIKEKLPG or derivatives of the fragments) rich in lysine residues, wherein the allele is formed by deficiency or insertion or replacement of one or more basic groups in the SEQ ID NO. 1 of the sequence table, the protein is formed in the way that an amino acid residue sequence shown in the SEQ ID NO. 2 of the sequence table are subjected to replacement or deficiency or addition through one or more amino acid residues, and the protein has the same activity as the amino acid residue sequence shown in the SEQ ID NO. 2 and is derived through the SEQ ID NO. 2. The obtained gene can be used for improving the salt resistance and heat resistance of plants and microorganisms.

Description

technical field [0001] The invention relates to the technical field of biogenetic engineering, in particular to the dehydrin gene of alfalfa bean and its application. Background technique [0002] Dehydrins belong to the LEAII family of late embryogenesis abundant proteins. Dehydrin was first found in seeds, and subsequent studies have shown that dehydrin is widely distributed in various tissues and organs of algae, yeast, nematodes, cyanobacteria and higher plants. Environmental factors such as drought, low temperature and high salinity that cause dehydration, as well as abscisic acid (ABA) can induce its expression in cells. Dehydrins contain at least one conserved lysine-rich motif K-fragment (EKKGIMDKIKEKLPG or its derivatives), which often form amphipathic (hydrophilic-lipophilic) helical structures. Some dehydrins usually also contain Y-fragment and S-fragment. The Y-fragment usually appears at the N-terminal of the dehydrin in 1-3 repeats (T / VDEYGNP), and the S frag...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N1/21C07K14/415C12R1/19
CPCY02A40/138
Inventor 王海庆沈迎芳窦全文陈志国
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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