Preparation method and application of urease activity inhibition yolk antibody

A technology of yolk antibody and urease activity, applied in the direction of anti-enzyme immunoglobulin and immunoglobulin from eggs, etc., can solve the problem of complex antigen components, difficulty in inhibiting yolk antibody, immunoglobulin specificity and antibody titer Not strong and advanced issues, to achieve the effect of high expression level, good protection effect, and cost reduction

Inactive Publication Date: 2016-03-02
李昕阳 +1
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Problems solved by technology

According to the regulations of the US Food and Drug Administration (FDA), drug-resistant plasmids cannot exist in live vaccines, and antibiotics cannot be used to maintain the stability of recombinant plasmid immune responses in humans and animals
Nie Zuoming and others use the extract of Helicobacter pylori antigen and HP urease that will be crushed by ultrasonic waves to immunize laying hens as a mixture of mixed (Urease) antigens to prepare anti-Hp IgY...

Method used

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Embodiment 1

[0022] A method for preparing egg yolk antibody inhibiting urease urease activity, the method comprising the following steps: performing gene mutation on the 1710bp DNA fragment of the Helicobacter pylori urease B subunit gene (see the appendix for the mutated sequence), and subcloning the mutated sequence into prokaryotic expression Vector, and form the fusion expression plasmid pET32a-ureB; Plasmid pET32a-ureB is transformed in Escherichia coli and form genetically engineered bacterium (the bacterial strain of this genetically engineered bacterium preservation microbial strains in China preservation manage committee Ordinary Microbiology Center, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, preservation dated October 15, 2015, preservation The number is CGMCCNO: 11509, and the classification is named as Escherichia coli), and Ni-NTA0.1~1.0mMIPTG is used to induce protein expression at...

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Abstract

The present invention belongs to the technical field of biology, and specifically discloses a preparation method and application of a urease activity inhibition yolk antibody. By use of gene recombination technology, helicobacter pylori urease (Urease) B subunit DNA fragment is cloned, and after base mutation, constructed and cloned on to a prokaryotic expression plasmid vector, genetic engineering strain BL21 (DE3) is transformed, IPTG is used for inducing expression of recombinant antigen protein urease B subunit, and a soluble form of expression can be successfully implemented; lysis of cells is performed, and urease B subunit recombinant protein can be isolated and purified; the purified recombinant protein urease B subunit as an antigen and an adjuvant are prepared into a microsphere vaccine for laying hen immunization, the yolk antibody in egg yolk can be extracted by pot group-type countercurrent extraction, and the yolk antibody titer and specificity can be detected by ELISA and other techniques. The obtained yolk antibody is added into milk for direct drinking for curing and prevention of acute and chronic gastritis, peptic ulcer and other stomach diseases caused by helicobacter pylori infection.

Description

technical field [0001] The invention belongs to the field of biotechnology and is suitable for preventing and treating gastric diseases such as acute and chronic gastritis and peptic ulcer caused by Helicobacter pylori infection, and specifically relates to a preparation method and application of egg yolk antibody for inhibiting urease activity. Background technique [0002] Helicobacter pylori (Hp) is closely related to the occurrence of gastric diseases such as acute and chronic gastritis, peptic ulcer, gastric cancer and gastric MALT lymphoma. In 1994, the World Health Organization classified it as a class of carcinogens. At present, the infection rate of Helicobacter pylori in developed countries reaches 30%, and the infection rate of Hp among people in developing countries is as high as 50%. After eradicating HP, it can not only accelerate the healing of gastric ulcer, but also significantly reduce the recurrence rate of gastritis. However, it is still very difficult ...

Claims

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Application Information

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IPC IPC(8): C07K16/40C07K16/02
Inventor 李昕阳张智
Owner 李昕阳
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