Protein GmGPA3 for regulating plant storage protein sorting as well as encoding gene and application thereof

A protein and plant technology, applied in the direction of plant genetic improvement, application, plant peptides, etc.

Active Publication Date: 2017-08-08
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular transport mechanism of this pathway has not been reported in soybean

Method used

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  • Protein GmGPA3 for regulating plant storage protein sorting as well as encoding gene and application thereof
  • Protein GmGPA3 for regulating plant storage protein sorting as well as encoding gene and application thereof
  • Protein GmGPA3 for regulating plant storage protein sorting as well as encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1, the cloning of soybean protein GmGPA3 gene

[0075] 1. Acquisition of cDNA

[0076] Total RNA was extracted from soybean Williams 82, and cDNA was synthesized by reverse transcription.

[0077] 2. PCR amplification

[0078] Using the cDNA obtained in step 1 as a template, the primer GmGPA3-cDNA-F / R was used for PCR amplification to obtain a PCR amplification product. The primer sequences are as follows:

[0079] GmGPA3-cDNA-F:5'-TTGAAACGAAACAAGAAGAATGC-3';

[0080] GmGPA3-cDNA-R: 5'-CGTAGGTTCAAACATGGCATAAA-3'.

[0081] 3. Sequencing

[0082] The PCR amplification products were sequenced. Sequencing results: PCR amplification obtained a band with a size of 1497bp, which had the nucleotides shown in sequence 1 in the sequence listing, the gene shown in sequence 1 was named GmGPA3 gene, and the protein encoded by the GmGPA3 gene was named GmGPA3. The amino acid sequence of the protein is sequence 2 in the sequence listing.

Embodiment 2

[0083] Example 2, the application of soybean protein GmGPA3 in regulating the sorting of plant storage proteins

[0084] 1. Preparation of transgenic GmGPA3 plants

[0085] 1. Preparation of recombinant vector

[0086] Between the XbaI recognition sequence of vector pCAMBIA1300-221, insert the DNA molecule shown in the 1st-1497th nucleotide of sequence 1 in the sequence table, and keep the other sequences of vector pCAMBIA1300-221 unchanged, obtain the recombinant vector, and put the The recombinant vector was named pCAMBIA1300-221-GmGPA3. The difference between pCAMBIA1300-221-GmGPA3 and pCAMBIA1300-221 is only that: pCAMBIA1300-221-GmGPA3 is obtained by inserting the DNA molecule shown in the 1st-1497th nucleotide of sequence 1 in the sequence table between the XbaI recognition sequence of pCAMBIA1300-221 recombinant vector. The pCAMBIA1300-221-GmGPA3 recombinant vector expresses the GmGPA3 protein shown in Sequence 2, and the expression of the GmGPA3 protein is initiated...

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Abstract

The invention discloses a protein GmGPA3 for regulating plant storage protein sorting as well as an encoding gene and application thereof. The GmGPA3 provided by the invention is shown as the following of a) or b) or c): the a) is a protein with the amino acid sequence being shown as sequence 2; the b) is a fused protein obtained by connecting a label to an N-terminal and / or a C-terminal of the protein shown by the sequence 2; the c) is a protein with the same function obtained by performing substitution and / or deletion and / or addition of one or multiple of amino acid residues on the amino acid sequence shown by the sequence 2. An experiment shows that the protein related to the plant storage protein sorting provided by the invention influences the sorting of a storage protein in a plant, and can be used for culturing transgenic plants with normal storage protein sorting.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a protein GmGPA3 regulating plant storage protein sorting, its coding gene and application. Background technique [0002] Soybean is an important oil and protein crop in my country. Soybean storage protein accounts for 70%-80% of soybean grain protein content, and is an important source of edible protein, as well as the source of amino acids and nitrogen required for seedling germination. Compared with other plants, the amino acid composition of soybean storage protein is relatively complete, especially rich in lysine. The health notice of the US Food and Drug Administration (FDA) shows that eating foods containing 25g of soy protein per day can help reduce the incidence of cardiovascular diseases. [0003] Soybean grain storage proteins are mainly synthesized in developing soybean cotyledons and deposited in the form of protein bodies. Taking 11S protein as an example...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8251
Inventor 邱丽娟魏中艳任玉龙
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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