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Protein GPA4 related to plant glutelin transportation and storage and encoding gene and application thereof

A related protein and gluten technology, which is applied to the plant gluten transport and storage related protein GPA4 and its encoding gene and application field, can solve the problems of unknown function and the like

Active Publication Date: 2017-02-22
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are also reports of GPA4 homologs in animals, such as human Got1A and Got1B, but their functions are unknown

Method used

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  • Protein GPA4 related to plant glutelin transportation and storage and encoding gene and application thereof
  • Protein GPA4 related to plant glutelin transportation and storage and encoding gene and application thereof
  • Protein GPA4 related to plant glutelin transportation and storage and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Discovery of related proteins and their coding genes for plant gluten sorting

[0048] 1. Phenotype analysis and genetic analysis of gpa4 mutant of rice gluten sorting

[0049] In 9311 60 The silty mutant gpa4-1 was selected from the Co radiation mutant library, and the silty mutant gpa4-2 was selected from the Kinmaze MNU-treated mutant library. The two mutants were allelic variants of the same gene. Compared with the wild type, the main feature of gpa4 is the grain quality and opacity (see figure 2 ). Scanning electron microscope analysis confirmed that the starch granules in gpa4-1 were loose (see figure 2 ), which may be the main cause of endosperm opacity. In addition, the SDS-PAGE analysis of the mature seeds of the mutant showed that the mutation of the gene resulted in a sharp increase in the 57-kDa gluten precursor, and the corresponding mature acidic and basic subunits decreased, along with the globulin content. Decreased, gliadin showed no significa...

Embodiment 2

[0089] Example 2. Obtaining and identification of transgenic plants

[0090] 1. Recombinant expression vector construction

[0091] Using rice Kinmaze cDNA as a template, PCR amplification was performed to obtain the coding region of GPA4 gene. The PCR primer sequence is as follows:

[0092] primer3: (The underline is the BamHI restriction site)

[0093] 5'CTGCAGGTCGAC GGATCC ATGGTTTCCTTCGAGATG 3'(SEQ ID NO. 6)

[0094] primer4: (the underline is the BamHI restriction site)

[0095] 5'TAGAATTCCCGG GGATCC CTACACTGGAACCCGTTT 3'(SEQ ID NO. 7)

[0096] The above-mentioned amplified product contains the coding region of the gene, and the PCR product is recovered and purified. The PCR product was cloned into the vector pCUbi1390 using Infusion Recombination Kit (Clontech) to construct pCUbi1390-GPA4. Recombination reaction system (2.5μL): PCR product 0.5μL (50-100ng), pCUbi1390 vector 1.0μL (30-50ng), 5×Infusionbuffer 2.0μL, Infusion enzyme mix 0.5μL. After a short centrifugation, the mi...

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PUM

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Abstract

The invention discloses protein GPA4 related to plant glutelin transportation and storage and an encoding gene and application thereof. The protein is protein formed by an amino acid sequence shown in the SEQ ID NO.1 or protein which is formed after the amino acid sequence shown in the SEQ ID NO.1 is subjected to substitution and / or deletion and / or addition of one or more amino acid residues, related to plant glutelin transportation and storage and derived from the SEQ ID NO.1. Plants with the normal mature glutelin content can be cultivated by transferring the encoding gene of the protein into plants with the low mature glutelin content. The protein and the encoding gene thereof can be applied to genetic improvement of the plants.

Description

Technical field [0001] The invention belongs to the field of genetic engineering, and relates to a plant gluten transport and storage related protein GPA4 and its coding gene and application. Background technique [0002] Higher plant seeds often accumulate a large amount of storage materials, which are used as a source of nutrients for seed germination and an important source of protein for humans and animal husbandry. Rice (Oryza sativa L.) is one of the most important food crops in the world, and more than 50% of the global population feed on rice as the staple food. Rice seeds accumulate a large amount of storage protein in the endosperm, which is the second largest substance in rice after starch, accounting for 8-10% of the dry weight of the grain, of which gluten can account for 60-80%, and prolamin is about It accounts for 10-20%, and globulin is about 5-10%. The content and composition of protein in rice directly affect the appearance, nutrition and eating quality of ri...

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 万建民王益华刘峰任玉龙鲍依群江玲刘喜
Owner NANJING AGRICULTURAL UNIVERSITY
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