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Stress-resistant protein and encoding gene thereof

A protein and stress resistance technology, applied in the field of molecular biology, can solve problems such as the lack of alfalfa dehydrin coding gene sequence, and achieve the effect of increasing economic benefits

Active Publication Date: 2018-01-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report related to the gene sequence encoding alfalfa dehydrin

Method used

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  • Stress-resistant protein and encoding gene thereof
  • Stress-resistant protein and encoding gene thereof
  • Stress-resistant protein and encoding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1, the cloning of alfalfa "WL-525HQ" MsDHN1 gene

[0068] 11. Acquisition of plant material

[0069] Take alfalfa "WL-525HQ" leaf tissue for RNA extraction;

[0070] 12. Extraction of RNA

[0071] The total RNA was extracted with the Plant Total RNA Extraction Kit of Beijing Quanshijin Biological Company, the integrity of the RNA was identified by formaldehyde denaturing gel electrophoresis, and then the purity and concentration of the RNA were determined on a spectrophotometer (Thermo Scientific NANODROP1000Spectrophotometer);

[0072] 13. Full-length cloning of genes

[0073] According to the EST sequence isolated from the results of the alfalfa "WL-525HQ" AL stress gene chip and the protein function annotation results, the core fragment of the alfalfa "WL-525HQ" MsDHN1 gene was obtained. The genome of Medicago truncatula has been sequenced, and the full-length cDNA was obtained by direct PCR according to the similarity between the two.

[0074] The ext...

Embodiment 2

[0075] Example 2. Sequence information and homology analysis of the alfalfa "WL-525HQ" MsDHN1 gene

[0076] The full-length open reading frame sequence of the alfalfa "WL-525HQ" MsDHN1 gene in the embodiment of the present invention is 669 bp, and the detailed sequence is shown in SEQ ID NO: 1. According to the open reading frame sequence, the amino acid sequence of the alfalfa "WL-525HQ" MsDHN1 protein was deduced, with a total of 222 amino acid residues, a molecular weight of 24.87kDa, and an isoelectric point (pI) of 5.63. For the detailed sequence, see SEQ ID NO: 4 sequence shown.

[0077] The open reading frame sequence of alfalfa "WL-525HQ" MsDHN and the amino acid sequence of its encoded protein were compiled in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDStranslations+PDB+SwissProt+Superdate+PIR using BLAST program Nucleotide and protein homology searches were carried out in the database, and it was found that it had 86.81% identity with the Medica...

Embodiment 3

[0078] Example 3. Expression difference of alfalfa "WL-525HQ" MsDHN1 gene at different stages of Al stress

[0079] 31. Material acquisition: samples were taken from leaves of alfalfa "WL-525HQ" Al stress materials at different stages (0h, 3h, 6h, 12h, 24h). Wrap the samples with aluminum platinum paper and put them into liquid nitrogen immediately, and then transfer them to -80°C ultra-low temperature refrigerator for storage until use;

[0080] 32. Extraction of RNA: using RNA prep pure plant total RNA extraction (Beijing Quanshijin); extraction of total RNA in different sample tissues of alfalfa "WL-525HQ";

[0081] 33. Determination of the integrity, purity and concentration of RNA: Use ordinary agarose gel electrophoresis (gel concentration 1.2%; 0.5×TBE electrophoresis buffer; 150v, 15min) to detect the integrity, and the maximum rRNA brightness in the electrophoresis band should be The brightness of the second rRNA is 1.5 to 2.0 times, otherwise it means that the rRNA ...

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Abstract

The invention relates to the technical field of molecular biology, in particular to isolated protein. The expression level of the protein in a plant body is related with the stress resistance of the plant, and the protein is protein (1) or protein (2); the protein (1) has an amino acid sequence shown in SEQ ID NO:4; the protein (2) has at least 80% of homology with the protein (1), and has the function of protein (1). The protein has the advantages that the stress resistance of plant, especially forage grass plant, is improved, and the economic benefit of the forage grass plant is increased.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to an antistress protein and its coding gene. Background technique [0002] Alfalfa (Medicago sativa.) is a very important forage grass species. It is a leguminous perennial herb with well-developed rhizomes and stolons. It has good palatability and high protein content. As a high-quality plant-based protein resource, it can greatly increase the output of crude protein compared with cereal crops due to its unique nitrogen utilization method. It is a solution to the development of the dairy industry. An important way and an effective method for the shortage of protein feed resources. [0003] Dehydrin belongs to the LEA-Ⅱ family and is a highly heat-stable and hydrophilic LEA protein (late embryogenesis abundant proteins) in plants. in the plant kingdom. It can be expressed in plants under adversity conditions such as drought, and its expression in adversity is closely r...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/54C12Q1/6895C12N15/11
Inventor 安渊周鹏吕爱敏刘星辰樊娜娜文武武
Owner SHANGHAI JIAO TONG UNIV
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