Preparation method of polyclonal antibody of wheat gliadin TaWG05

A wheat prolamin and polyclonal antibody technology, which can be used in botany equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., and can solve problems such as wheat allergy

Inactive Publication Date: 2017-02-22
HENAN ACAD OF AGRI SCI XIAOMAI INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there is a serious problem in wheat application, namely wheat allergy

Method used

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  • Preparation method of polyclonal antibody of wheat gliadin TaWG05
  • Preparation method of polyclonal antibody of wheat gliadin TaWG05
  • Preparation method of polyclonal antibody of wheat gliadin TaWG05

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The wheat γ-glamin TaWG05 polyclonal antibody provided in this application is prepared through the following steps.

[0058] (1) Genomic DNA of wheat variety Zhengmai 004 was extracted;

[0059] Genomic DNA of wheat variety Zhengmai 004 was extracted by using the Plant Genomic DNA Extraction Kit of Tiangen Company and referring to its instructions.

[0060] (2) Design primers: when designing primer sequences, based on the prediction of the bioinformatics software SignalP 4.1 server, the 22 amino acid residues at the N-terminal of wheat γ-gliadin TaWG05 are transit peptides (the prediction results are as follows: figure 1 shown), therefore, when amplifying the target gene corresponding to wheat γ-gliadin TaWG05, the PCR amplification primers for removing its transit peptide were designed as follows:

[0061] Upstream primer: TCGACCCTCGCATCCATGACCAA,

[0062] Downstream primer: TCATTGTCCACCATTGTAGGCG;

[0063] Using the genomic DNA extracted in step (1) as a template, ...

Embodiment 2

[0120] Using the polyclonal antibody against γ-prolamin TaWG05 prepared in Example 1, it is possible to effectively detect whether other wheat varieties (or wheat-containing protein products) contain γ-prolamin TaWG05, so as to detect γ-prolamin TaWG05. Protein TaWG05 allergic people provide effective health guidance, and also provide guidance for the improvement of wheat varieties. Specifically, this example takes some wheat varieties (Xinmai 26, Zhengmai 366, Zhengmai 9023, Aikang 58, Zhengmai 004, Yumai 18, Yanzhan 4110) as examples, and whether they contain γ-alcohol The solubilized protein TaWG05 protein has been detected, and the detection results can lay the foundation for wheat improvement or anti-allergen detection. The specific experimental process is as follows.

[0121] (1) Extract wheat seed protein, the specific steps are as follows:

[0122] (A) Add 200 μL total protein extraction buffer into a 1.5ml centrifuge tube and put it on ice for later use;

[0123] T...

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Abstract

The invention belongs to the technical field of biological technology, and particularly relates to a preparation method of polyclonal antibody of wheat gliadin TaWG05. The method comprises the steps of extracting the genome DNA of the wheat variety of Zhengmai 004, designing primer, conducting the PCR amplification, and constructing recombinant plasmid pET-32A-TaWG05, conducting prokaryotic expression, conducting immunization, and preparing the polyclonal antibody and the like. The application of polyclonal antibody can establish the reliable and quantitative allergen test method, thus providing the guarantee for the consistency of food labeling and the safety of consumers. Moreover the polyclonal antibody is used to test and authenticate the expression and distribution status of the wheat gliadin TaWG05 in other wheat varieties, thus laying down the theoretical foundation for the modification of wheat varieties.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of wheat prolamin TaWG05 polyclonal antibody. Background technique [0002] Wheat is one of the important food crops in my country. Wheat flour can be used as the main raw material to make many foods, such as bread, fried dough sticks, pancakes, dumplings, noodles, etc. It plays an important role in people's food structure and food consumption. The gliadin in wheat has the function of maintaining the stomach, can form a protective film on the surface, effectively prevent oxidation, can improve the purity of gastric mucosal protein, and can prevent gastric acid erosion to a certain extent, and gliadin has anti-inflammatory effect, is the Increased resistance. [0003] However, there is a serious problem in wheat application, namely wheat allergy. Wheat allergy refers to the adverse reaction of certain people to some proteins in flour. Since wheat an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/16C07K16/06C07K14/415C12N15/29C12N15/70G01N33/68
CPCC07K14/415C07K16/06C07K16/16C12N15/70C12N2800/101G01N33/68G01N2333/415
Inventor 秦毛毛周正富刘聪聪汪庆昌雷振生吴政卿常阳王亚欢晁岳恩王美芳何盛莲李文旭杨攀刘加平徐福新李巍张琨
Owner HENAN ACAD OF AGRI SCI XIAOMAI INST
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