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Sheep brucella M5 strain ribosome protein L7/L12 gene, encoding protein and uses thereof

A Brucella and ribosomal protein technology, applied in the field of genetic engineering, can solve problems such as strong toxic reactions, health hazards in animal husbandry, susceptibility to Brucella melis, etc.

Inactive Publication Date: 2008-05-14
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there are many deficiencies in the existing vaccines, and the crowd is the most susceptible to Brucella melis and has the strongest toxic reaction, the infection rate of Brucellosis has been rising in a straight line in recent years, which has a great impact on the development of animal husbandry and human beings. serious harm to the health of

Method used

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  • Sheep brucella M5 strain ribosome protein L7/L12 gene, encoding protein and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Obtaining and identification of Brucella ovis M5 ribosomal protein L7 / L12 gene

[0015] 1. Culture of Brucella melis M5 strain

[0016] First, resuscitate Brucella melis M5 strain freeze-dried bacteria (CMCC55009, purchased from China Institute for the Control of Pharmaceutical and Biological Products) with Brucella broth medium, and Brucella broth medium (purchased from China Import and Export Commodity Inspection Technology Institute) ) on a solid slant for streak culture at 37°C, 5% CO 2 Cultivate under the same conditions for 10 days, after Brucella grows on the solid slope to form a single colony visible to the naked eye, carefully scrape a single colony with an inoculation loop, inoculate it into liquid Brucella broth medium, and culture it in a water bath shaker at 37°C After 24 hours, a bacterial solution with a suitable concentration was obtained.

[0017] 2. Extraction of Bacterial Whole Genomic DNA

[0018] Use the cultured bacterial solution wit...

Embodiment 2

[0024] Example 2 Brucella melis M5 strain L7 / L12 gene and (control strain) 16M strain

[0025] Effect comparison

[0026] Objective: To investigate the immune effect of ribosomal protein L7 / L12 gene obtained from Brucella melis M5 strain (CMCC 55009) Immunity effects were compared.

[0027] 1. Plasmid construction

[0028]The L7 / L12 gene (L7 / L12 5 ) and the 16M strain L7 / L12 gene (L7 / L12 16 , GenBank L27819) were respectively constructed on the eukaryotic expression vector pcDNA3.1(+) to obtain pcDNA3.1(+)-L7 / L125 and pcDNA3.1(+)-L7 / L12 16 Two plasmids. The above two plasmids constructed above were respectively transfected into Escherichia coli E. coli DH5α, and screened in the presence of 100 ug / ml ampicillin. The positive clones obtained by screening were expanded and cultured, and a large amount of recombinant plasmids were extracted, and the plasmids were diluted to 1000ug / ml with 0.01MPBS (pH7.2) buffer.

[0029] 2. Experimental animals and reagents

[0030] 1. Ex...

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Abstract

The invention provides a gene of strain ribosome protein L7 / L12 of brucellosis M5 of sheep type and encoded protein and application thereof. The encoded protein of the gene of the invention has an amino acid sequence shown in SEQ ID No.2. The gene of the invention can be used for preparing gene vaccine. For example, the siRNA or shRNA or microRNA can be synthesized and used for gene silencing by the sequence of the invention. By preparing the expression protein of the gene of the invention, the recombinant protein vaccine can be researched, or a test kit can be developed, which provides an effective solving way for the brucellosis.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to the gene of ribosomal protein L7 / L12 of Brucella melitensis M5 strain, its coded protein and application. Background technique [0002] Brucellosis is a zoonotic infectious disease caused by Brucella. Brucella is a mammalian parasite that is highly infectious and pathogenic to humans and mammals. The genus has seven biological species: sheep, cattle, pigs, dogs, wood rats, marine mammals and sheep. The first four species are mainly popular in China. Livestock such as sheep, cattle, and pigs are most susceptible to infection, and sheep are the most common. Because Brucella is a facultative intracellular parasite, it not only parasitizes the mononuclear-macrophage system, but also reproduces in placental villous epithelial cells, renal parenchymal cells, testicular fibroblasts, and embryonic cells, causing reproductive Inflammation of organs and membranes, abortion, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/23A61K48/00C12Q1/68A61K39/10G01N33/68
Inventor 石艳春郑源强旭日干
Owner INNER MONGOLIA UNIVERSITY
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