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Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof

A gluten subunit, capillary electrophoresis technology, applied in cleaning methods and utensils, chemical instruments and methods, measuring devices, etc., to achieve the effects of high repeatability, high speed, and low dosage

Inactive Publication Date: 2012-01-18
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main task is to solve the identification of low molecular weight glutenin subunits stored in wheat seeds, and to screen out the optimal electrophoresis parameters for capillary electrophoresis

Method used

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  • Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof
  • Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof
  • Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1, Construction of Wheat Low Molecular Weight Glutenin Subunit Standard CE Fingerprint

[0047] (1) Sampling and sample preparation

[0048] Take a single seed of wheat "Chinese spring" (Chinese spring seeds are bred and stored in the Genetic and Bioengineering Laboratory of Capital Normal University), remove the embryo end with a blade, accurately weigh 200 mg, and smash it into fine pieces with sulfuric acid paper and a small hammer. Put the powder into a 1.5ml centrifuge tube; add 1ml of ethanol with a volume percentage of 70%, vortex for 30-60min, centrifuge at 10000rpm for 10min, and remove the supernatant. Add 1ml of 55% isopropanol by volume, bath in 65°C water for 30min, centrifuge at 10000g for 10min, remove the supernatant and blot dry with filter paper, repeat 3 times. Add 0.1ml of 1% DTT and 0.1ml of solution A, wherein the composition of solution A is: isopropanol: 1M Tris-HCl (pH6.8): double distilled water = 5:2:3, vortex to mix , 65°C water bat...

Embodiment 2

[0077] Embodiment 2, using the standard map constructed in embodiment 1 to identify wheat varieties

[0078] (1) Experimental operation: Take the seeds of 6 different varieties of wheat, including Jimai 20, Zhongyou 9507, Zhoumai 16, Xiaoyan 6, Jingdong 8, and Jing 411 (provided by the Institute of Crop Science, Chinese Academy of Agricultural Sciences) , Sampling and sample preparation, capillary electrophoresis, and construction of a standard map are the same as in Example 1.

[0079] (2) CE pattern analysis: According to the differences in migration time, peak height, peak area and other parameters of low molecular weight glutenin subunit components of different wheat varieties, their electrophoretic composition characteristics were analyzed. Repeat the electrophoresis 3 times for each wheat variety, calculate the relative standard deviation (RSD) of peak migration time, peak height, and peak area, and obtain the average value and relative standard deviation of the water-so...

Embodiment 3

[0082] Example 3, using the standard map constructed in Example 1 to identify different LMW-GS alleles of wheat varieties

[0083](1) Experimental operation: Take the seeds of different varieties of wheat such as Jimai 20, Zhongyou 9507, Zhoumai 16, Xiaoyan 6, Jingdong 8, Jing 411 (provided by the Institute of Crop Science, Chinese Academy of Agricultural Sciences), and take samples. And sample preparation, capillary electrophoresis, map analysis are equal to embodiment 1 and embodiment 2.

[0084] (2) CE pattern analysis: According to the differences in the migration time, peak height, peak area and other parameters of the low molecular weight glutenin subunit components of different wheat varieties, their electrophoretic composition characteristics were analyzed. Repeat the electrophoresis 3 times for each variety, calculate the relative standard deviation (RSD) of peak migration time, peak height, and peak area, and obtain the average value and relative standard deviation o...

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Abstract

The invention discloses a capillary electrophoresis method for identifying a low molecular weight glutelin subunit of wheat. Through the method, a standard capillary electrophoresis pattern of the low molecular weight glutelin subunit of the wheat can be established, and further, a set of complete technical systems for identifying the high-quality subunit of the wheat is formed. According to protein peak migration time and peak area identified variety and germplasm, screened high-quality subunits and allelic genes, the rapid improvement on the quality of the wheat is realized.

Description

technical field [0001] The invention relates to a rapid separation and identification method for wheat low molecular weight glutenin subunits, in particular to the identification of wheat low molecular weight glutenin subunit alleles and varieties (lines) with different quality characteristics by capillary electrophoresis (CE) technology Methods. Background technique [0002] Wheat is an important food crop for humans and an important source of protein for humans. The protein stored in its grain is mainly composed of gliadin and glutenin. The composition and content of these two proteins determine the quality of wheat to a large extent. It is an important genetic marker for wheat variety identification and quality improvement. The molecular weight of glutenin in the storage protein is relatively large, which is an important factor affecting the viscoelasticity of wheat gluten. According to the difference in mobility in SDS-PAGE under reducing conditions, it can be divided ...

Claims

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Application Information

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IPC IPC(8): G01N27/447G01N1/28B08B9/02
Inventor 晏月明李婧
Owner CAPITAL NORMAL UNIVERSITY
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