Abortus Brucella vaccine strain S19 marked recombinant strain and application thereof
A technology of Brucella abortus and recombinant strains, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of strong virulence, difficult operation, hindering the eradication and purification of animal brucellosis, and achieve Low toxicity, promotes eradication and purification effects
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Embodiment 1
[0033] Embodiment 1, the construction of the recombinant bacterial strain of inactivating glucosyltransferase gene
[0034] According to the glucosyltransferase gene (GenBank AccessionNumber: BAbS19_I09300) in the Brucella abortus genome, two pairs of primers were designed, one pair of primers was used to amplify homology arm DA6-1, and the other pair of primers was used to amplify DA6- 2. Homology arm DA6-1 and homology arm DA6-2 are connected to form sequence DA6, and sequence DA6 is inserted into pEX18AP plasmid to obtain recombinant vector pEX18AP-DA6. Homologous recombination between recombinant vector pEX18AP-DA6 and Brucella abortus strain S19 can obtain a recombinant strain with inactivated glucosyltransferase gene.
[0035] 1. Construction of recombinant vector pEX18AP-DA6
[0036] For a schematic diagram of the construction process, see Figure 5 .
[0037] 1. Extraction of Brucella genome
[0038] Take 15 μl of inactivated Brucella abortus strain S19, add 1ml T...
Embodiment 2
[0080] Embodiment 2, the application of recombinant bacterial strain RB6
[0081] Experimental animals: 4-6 weeks old SPF grade Balb / C female mice, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number SCXK (Beijing) 2007-0001.
[0082] 1. Toxicity identification
[0083] The mice were randomly divided into four groups, 30 in each group. The specific vaccination plan is as follows:
[0084] The first group (test group): inoculate the recombinant bacterial strain RB6 that embodiment 1 prepares, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.
[0085] The second group (control group 1): inoculate Brucella abortus bacterial strain 2308, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.
[0086] The third group (control group 2): inoculate Brucella abortus bacterial strain S19, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.
[0087] The fourth group ...
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