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Specific augmentation primer and its method of labelling wheat macromolecular glutelin subunit

A technology of gluten subunit and amplification primer, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, sugar derivatives, etc. Accuracy and other issues, to achieve good promotion and application prospects, fast and obvious effects, and simple structure.

Inactive Publication Date: 2003-04-16
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that it is not accurate enough, and the second is that the detection and screening must wait until the next generation of seeds are harvested, which greatly affects the efficiency of breeding
However, the design of subunit-specific primers is quite difficult due to the highly conserved nature of the genes controlling high-molecular-weight glutenins in wheat
Therefore, there are few molecular markers currently available to identify wheat high-molecular glutenin subunits, and there is no Dx t The existence of specific primers for the 1.5 subunit limits the applicability of this method

Method used

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  • Specific augmentation primer and its method of labelling wheat macromolecular glutelin subunit
  • Specific augmentation primer and its method of labelling wheat macromolecular glutelin subunit
  • Specific augmentation primer and its method of labelling wheat macromolecular glutelin subunit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Using subunit-specific primers to identify whether the high-molecular-weight glutenin encoded by the D genome in different strains of Triticum japonicus and wheat contains the high-molecular-weight glutenin subunit Dx t 1.5.

[0037] The reaction system used is as follows:

[0038] PCR reaction system (20ul): 1×PCR buffer

[0039] 2mM Mg

[0040] 0.2mM of each dNTP,

[0041] 1U Ex-Taq DNA polymerase

[0042] 250ng AS-PCR primer (according to claim 1)

[0043] DNA template (appropriate amount)

[0044] Add water to 20ul

[0045] The PCR program is:

[0046] 6) Pre-denaturation at 94°C for 4 minutes

[0047] 7) Denature at 94°C for 1 minute

[0048] 8) Anneal at 65°C for 30 seconds

[0049] 9) Extend at 72°C for 1 minute

[0050] 10) Extend at 72°C f...

Embodiment 2

[0057] Using subunit-specific primers to identify whether the high-molecular-weight glutenin encoded by the D genome in a recombinant inbred wheat plant contains the high-molecular-weight glutenin subunit Dx t 1.5.

[0058] The reaction system used is as follows:

[0059] PCR reaction system (20ul): 1×PCR buffer

[0060] 2mM Mg

[0061] 0.2mM of each dNTP,

[0062] 1U Ex-Taq DNA polymerase

[0063] 250ng AS-PCR primer (according to claim 1)

[0064] DNA template (appropriate amount)

[0065] Add water to 20ul

[0066] The PCR program is:

[0067] 11) Pre-denaturation at 94°C for 4 minutes

[0068] 12) Denature at 94°C for 1 minute

[0069] 13) Annealing at 71°C for 55 seconds

[0070] 14) Extend at 72°C for 1 minute

[0071] 15) Extend at 72°C ...

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Abstract

A kind of 3 specific amplimers and a method for labelling high-molecular glutelin subunit with it are disclosed. Based on the polymerase chain amplification reaction, the difference in DNA sequences of different high-molecular glutelin subunits and the high speed and efficiency of PCR are used to correctly detect if wheat or Jiejie wheat contains the high-molecular glutelin subunit Dxt 1.5 from the Jiejie wheat, or the high-molecular glutelin subunit Dx2 or Dx5 of wheat, or the high-molecular glutelin subunit Dxt2 or Dxt5 of Jiejie wheat. Its advantages are high speed, and high effect.

Description

technical field [0001] The invention relates to a specific amplification primer and a method for marking wheat macromolecule glutenin subunit. technical background [0002] High molecular weight glutenin is an important storage protein in common wheat (Triticum aestivum) seeds, and its composition and expression level have a great influence on the processing quality of wheat. High molecular weight glutenin in wheat is encoded by a pair of closely linked genes located in the long arm of the first homologous transformation group, encoding two high molecular weight glutenin subunits with different molecular weights, namely: x subunit and y subunit. Due to gene silencing, usually only 3-5 high-molecular-weight glutenin subunits are expressed in hexaploid common wheat. It is currently believed that the high molecular weight glutenin subunit encoded by the D genome has the greatest impact on wheat quality. The D genome of wheat itself has low genetic diversity due to geographica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/00C12N15/10C12P19/34C12Q1/68
Inventor 卢宝荣陆春明杨武云
Owner FUDAN UNIV
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