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Gene of coded recombined helicobacter pylori serine protease, egg yolk antibody and application

A technology of Helicobacter pylori and myceline, applied in application, antibody, genetic engineering and other directions, can solve problems such as danger

Active Publication Date: 2018-09-14
北京宇诚健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, as these researchers discovered, the stage after the first is the more dangerous

Method used

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  • Gene of coded recombined helicobacter pylori serine protease, egg yolk antibody and application
  • Gene of coded recombined helicobacter pylori serine protease, egg yolk antibody and application
  • Gene of coded recombined helicobacter pylori serine protease, egg yolk antibody and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Cloning of embodiment 1 recombinant Helicobacter pylori (HP) serine protease (HtrA)

[0057] (1) Retrieve the amino acid sequence of Helicobacter pylori (HP) serine protease (HtrA) from the PubMed protein database. The selected number is GenBank: AAD05980.1. The amino acid sequence is as follows, and its codons are optimized:

[0058]

[0059] (2) Select pET-22b as the expression vector, BL21(DE3) as the expression host, and 5'Nde I and 3'SalI as the enzyme cutting sites, and entrust Sangon Bioengineering (Shanghai) Co., Ltd. to synthesize the full sequence of the gene And clone, obtain genetically engineered bacterium HPHtrA, wherein, the DNA sequence after codon optimization is as follows:

[0060]

[0061]

[0062] (3) The genetically engineered bacterium HPHtrA was inoculated in LB liquid medium containing Amp (final concentration: 100 μg / mL), cultured overnight at 37°C with shaking, and transferred at an inoculum size of 1% by volume the next day, and cult...

Embodiment 2

[0064] Example 2 Preparation of egg yolk antibody inhibiting Helicobacter pylori serine protease

[0065] (1) The recombinant Helicobacter pylori (HP) serine protease (HtrA) that embodiment 1 makes is mixed with the solution that protein concentration is 1mg / mL as immune antigen with physiological saline; Get 20 week-old laying hens 4, put After the immune antigen was mixed with Freund's complete adjuvant in equal volume and fully emulsified, the dose of 0.5 mL / hen was injected into the hen's wings or breast muscle at multiple points; the first booster immunization was carried out 2 weeks after the first immunization, using Freund's After the complete adjuvant and immune antigen were mixed in equal volumes and fully emulsified, the dose of 0.5 mL / hen was injected into the wings or breast muscles of hens at multiple points; thereafter, booster immunization was given every 15 days, and a total of 4 booster immunizations were given. Collect eggs after booster immunization, collec...

Embodiment 3

[0068] Example 3 Preparation of Egg Yolk Antibody Inhibiting Helicobacter Pylori Serine Protease

[0069] (1) The recombinant Helicobacter pylori (HP) serine protease (HtrA) that embodiment 1 makes is mixed with the solution that protein concentration is 0.4mg / mL as immune antigen with physiological saline; Get 20 week-old laying hens 4, After mixing the immune antigen with Freund's complete adjuvant in equal volume and fully emulsifying it, inject it at multiple points on the hen's wings or breast muscle at a dose of 0.5 mL / hen; carry out the first booster immunization 2 weeks after the first immunization, using Freund's After the incomplete adjuvant and the immune antigen were mixed in equal volumes and fully emulsified, the dose of 0.5 mL / hen was injected into the wings or breast muscle of the hen at multiple points; thereafter, booster immunization was given every 15 days, and a total of 5 booster immunizations were given. Collect eggs after the first booster immunization,...

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Abstract

The invention belongs to the technical field of biology, and specifically relates to a gene of coded recombined helicobacter pylori serine protease, an egg yolk antibody and application. The nucleotide sequence of the gene of the coded recombined helicobacter pylori serine protease is shown in SEQ ID NO.1. The invention further provides an egg yolk antibody inhibiting helicobacter pylori serine protease. The antibody is prepared through the following steps of performing gene synthesis to obtain the gene of the coded recombined serine protease; performing prokaryotic expression on the gene, purifying, acquiring recombined serine protease, using the recombined serine protease as an immunogen to immunize hens, collecting eggs laid by immunized hens, extracting the egg yolk antibody in the egg, purifying, and obtaining the egg yolk antibody inhibiting helicobacter pylori serine protease. The egg yolk antibody can be used as a food additive or added into healthcare products, or can be prepared into drugs preventing or treating gastric ulcer and gastric cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a gene encoding recombinant Helicobacter pylori serine protease, egg yolk antibody and application. Background technique [0002] Helicobacter pylori (Helicobacter pylori) was discovered by scientists Barry Marshall and Robin Warren, which can infect stomach tissue and cause ulcers, and the two researchers also won the 2005 Nobel Prize in Physiology and medical awards. [0003] Helicobacter pylori, the causative agent of gastric ulcer and gastric cancer. It is estimated that half of the world's population is infected with Helicobacter pylori. Many infected people develop symptoms of gastritis, and one in five infected people develops a stomach ulcer at some point in their lives, a rapidly progressive and fatal disease in many cases. Gastritis and stomach ulcers can be treated with antibiotics; these treatments often require the use of two or three different types of ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C12N15/57C12N15/70C12N1/21C07K16/40C07K16/02A61K39/395A61P35/00C12R1/19
CPCA61K2039/505A61P35/00C07K16/40C07K2317/11C12N9/6408C12Y304/21106
Inventor 赵肃清肖益热郑希王甜甜陈莉莉贺攀
Owner 北京宇诚健康科技有限公司
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