Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat

A technology of capillary electrophoresis and gluten subunit, which is applied in the directions of measuring device, material separation, analysis material, etc., can solve the problems of high analysis cost, high cost, labor and time consuming, etc., and achieves high resolution, low sample consumption, Easy-to-apply effects

Inactive Publication Date: 2005-01-26
CAPITAL NORMAL UNIVERSITY
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, although the improvement of wheat quality through genetic engineering and molecular marker-assisted selection has shown great application prospects, these methods are often expensive, high in analysis costs, labor-intensive and time-consuming, and the genetic engineering and The molecular marker-assisted selection method is still far from practical application, and it is still difficult to meet the requirements of quality breeding work

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat
  • Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat
  • Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1. Material sampling

[0051] The seeds of the wheat variety Hope were used as test materials, and 10-15 mg of the endosperm-free end of the seeds were randomly selected for gluten extraction and capillary electrophoresis.

[0052] 2. Sample Preparation

[0053] High molecular weight glutenin subunit (HMW-GS) extraction method:

[0054] Alternative reagents:

[0055] A. 50% n-propanol

[0056] B. 50% n-propanol + 1% DTT

[0057] C. Acetone (analytical pure)

[0058] D. 20% acetonitrile + 0.1% trifluoroacetic acid (TFA)

[0059] Operation method:

[0060] (1) Take a single seed, cut off the endosperm-free end (about 10-15mg) with a blade, weigh it accurately and use sulfur

[0061] Crush the acid paper and a small hammer into fine powder, and put it into a 1.5ml centrifuge tube;

[0062] (2) Add 1ml of 50% n-propanol, extract in a water bath at 45°C for 10 minutes, and centrifuge at 10000rpm for 5 minutes,

[0063] Remove the supernatant;

[0064] (3) Repeat th...

Embodiment 2

[0116] 1. Material sampling

[0117] The seeds of wheat varieties China Spring, Hope and Durum wheat were used as test materials respectively, and 10-15 mg of the endosperm-free end of the seeds were randomly selected for gluten extraction and capillary electrophoresis.

[0118] 2. Sample Preparation

[0119] High molecular weight glutenin subunit (HMW-GS) extraction method:

[0120] The selected spare reagents and operation method are the same as in Example 1, and the obtained HMW-GS precipitate can be fully dissolved by adding 20% ​​acetonitrile+0.1% w / v trifluoroacetic acid (TFA) in proportion (w / v: 1 mg+10 μl) , Capillary electrophoresis analysis can be performed after centrifugation; the precipitate can also be stored at -20°C, and dissolved when needed for capillary electrophoresis analysis.

[0121] 3. High performance capillary electrophoresis (HPCE)

[0122] equipment

[0123] High-efficiency capillary electrophoresis system (equipped with software for system oper...

Embodiment 3

[0165] 1. Material sampling

[0166] The seeds of club wheat Club20 and bread wheat variety Kontrast were used as test materials respectively, and 10-15 mg of the endosperm-free end of the seeds were randomly selected for gluten extraction and capillary electrophoresis. Then, the samples prepared by mixing the high molecular weight glutenin subunits extracted from the seeds of club wheat Club20 and Kontrast in a ratio of 1:1 were used as test materials.

[0167] 2. Sample Preparation

[0168] High molecular weight glutenin subunit (HMW-GS) extraction method:

[0169] The selected spare reagents and operation method are the same as in Example 1, and the obtained HMW-GS precipitate can be fully dissolved by adding 20% ​​acetonitrile+0.1% w / v trifluoroacetic acid (TFA) in proportion (w / v: 1 mg+10 μl) , Capillary electrophoresis analysis can be performed after centrifugation; the precipitate can also be stored at -20°C, and dissolved when needed for capillary electrophoresis ana...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
The inside diameter ofaaaaaaaaaa
Lengthaaaaaaaaaa
The inside diameter ofaaaaaaaaaa
Login to View More

Abstract

This invention relates to a determination method for wheat high molecular weight glutelin subunit, especially a method through technique of acid capillary electrophoresis. The key emphasis in current wheat breeding work is to improve the HMW-GS component, especially to improve the frequency of excellent high molecular weight glutelin subunit or subunit pairs The combination of current usual breeding method with label assistance selection techniques is the main means for wheat breeding improvement, wherein the rapid, microscale and accurate determination of excellent protein subunit for generation in the early hybrid time is the key to improve the breeding efficiency. The main task of this invention is to provide the buffer type and additives and condition parameter of acid capillary electrophoresis and clean-up procedures of capillary.

Description

technical field [0001] The invention relates to a method for identifying wheat high-molecular-weight glutenin subunits, in particular to a method for identifying wheat high-molecular-weight glutenin subunits through acid capillary electrophoresis (A-CE) technology. Background technique [0002] Wheat seed storage proteins are mainly composed of gliadins and glutenins, which are highly heterogeneous and complex in composition. The molecular weight of gliadin is between 30,000-80,000 Daltons. Under the condition of acidic gel electrophoresis, a variety can separate 15-30 components, which can be divided into α, β, γ and ω according to their relative mobility. Four gliadins accounted for 25%, 30%, 30% and 15% of the total amount respectively. Glutenin includes high molecular weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW-GS), which account for about 10% and 30% of the total protein content of endosperm respectively. [0003] A large number of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/06G01N30/28G01N30/38
Inventor 晏月明余建中姜怡安学丽胡英考
Owner CAPITAL NORMAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com