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Preparation process for gene vaccines containing UreB and CagA protein of helicobacter pylori

A Helicobacter pylori and gene vaccine technology, applied in the field of biomedicine, can solve the problem of low immune level and achieve the effect of solving low immune level

Inactive Publication Date: 2012-09-26
CHENGDU KANGHUA BIOLOGICAL PROD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The present invention overcomes the deficiency of low immune level of the existing single Helicobacter pylori gene vaccine, and provides a preparation method of a gene vaccine combined with Helicobacter pylori UreB and CagA proteins to improve immunity and avoid Helicobacter pylori infection Purpose

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  • Preparation process for gene vaccines containing UreB and CagA protein of helicobacter pylori
  • Preparation process for gene vaccines containing UreB and CagA protein of helicobacter pylori
  • Preparation process for gene vaccines containing UreB and CagA protein of helicobacter pylori

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Embodiment Construction

[0022] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail through the following examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0023] 1 Construction and identification of fusion gene ureB-cagA:

[0024] Recombinant plasmids pUC-ureB and pUC-cagA were cut forward with EcoR Ⅰ and Pst Ⅰ ​​respectively, the target fragments were recovered by tapping rubber, ligated and transformed, the resistant clones were picked and amplified, and the plasmids were extracted to obtain pUC-ureB-cagA. All identification.

[0025] 1.1 Digestion of recombinant plasmids

[0026]

[0027]

[0028] Digest for 1 hour at 37°C, stop the reaction with 2ul 10xLoadding Buffer, and electrophoresis on 1.0% agarose gel

[0029] 1.2 Construction and identification of recombinant plasmi...

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Abstract

The invention provides a preparation process for gene vaccines containing UreB and CagA protein of helicobacter pylori. The preparation process includes: establishing recombinant plasmid pIRES-UreB-CagA by recombinant plasmids pUC-UreB and pUC-CagA according to the genetic engineering technology, transforming, screening, amplifying and determining; chemically transforming attenuated Salmonella typhimurium LB500 to perform methylation; screening ampicillin resistant bacterial colony; amplifying selectively; extracting a small quantity of plasmids by the alkaline lysis method; obtaining definitive host bacteria SL7207 by further pulse cell transfection; picking resistant colone to amplify the same in LB culture solution containing 100ug / ml ampicillin to obtain protein; and finally obtaining helicobacter pylori vaccines by purifying the protein. The novel vaccines containing UreB and CagA protein of helicobacter pylori has immunization effects of both UreB and CagA protein, so that immunization efficacy can be improved to a high level.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of biomedicine, in particular to a method for preparing a gene vaccine containing Helicobacter pylori UreB and CagA proteins. Background technique [0002] It has been confirmed that Helicobacter pylori (H. Pylori, Hp) is closely related to some diseases of the upper gastrointestinal tract, including gastric mucosa-associated lymphoid tissue lymphoma (MALT), chronic gastritis, peptic ulcer, and gastric cancer. The whole Helicobacter pylori bacteria has a very complex antigen composition, which is not easy to preserve, has a long production cycle, low yield, and is easily contaminated. With the development of molecular biology, people have quickly invested in the research of various special genes of Hp, and have successfully cloned more than ten different genomes or genes of Hp, including: chemokines cheA and cheY, cytotoxin-related gene cagA And cagC, flagellin gene flaA and flaB, ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/02A61P31/04C12N15/74C12R1/42C12R1/01
CPCY02A50/30
Inventor 侯文礼蔡勇钟泽荣冯晓
Owner CHENGDU KANGHUA BIOLOGICAL PROD
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