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.
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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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