Attenuated Salmonella choleraesuis s.choleraesuis S1210 and its preparation method and application
A technology of Salmonella cholerae, CCTCCM2015632, applied in biochemical equipment and methods, microorganism-based methods, bacteria, etc., can solve problems such as not being very clear, and achieve excellent detoxification effect.
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experiment example 1
[0052] Phenotypic Characterization of Experimental Example 1 Gene Deletion Strain
[0053] We will compare and observe the lipopolysaccharides of the strain obtained in Example 1 and the wild strain, which are important immunogens or virulence factors of Salmonella choleraesuis.
[0054] Lipopolysaccharide extraction and silver staining observation: extract a small amount of LPS for silver staining observation, the specific steps are: take 2-3mL each of the bacterial strain obtained in Example 1 and the wild strain cultivated overnight, centrifuge at 1,2000×g for 1min, Collect the cells; wash the cells with PBS or ultrapure water 2 to 3 times; take 150 μL of lysis buffer (1 mL 0.5M Tris-cl PH=6.8, 0.8 mL glycerol, 1.6 mL 10% SDS, 0.4 mL β-mercaptoethanol, 4.2 mL Ultrapure water), mix the cells and cook in boiling water for 10 minutes; cool the sample to room temperature, centrifuge at 1,2000×g, 4°C for 10 minutes; take 10 μL of the supernatant and add it to 90 μL loading buffe...
experiment example 2
[0056] Experimental example 2 Determination of attenuation effect of Salmonella choleraesuis wbaD gene deletion strain
[0057] In order to determine the attenuation effect of the constructed gene deletion strain C3545ΔwbaD, the virulence changes of the gene deletion strain and the parental strain on mice were compared by oral administration. In this experiment, six-week-old BALB / c female mice were purchased, fed for one week to adapt to the environment, and then challenged by oral routes. There were 3 mice in each group, a total of eight groups, of which six groups were mutant strains, one group was a blank group that took BSG orally, and one group was oral administration of BSG. It is the positive control group of oral wild strain C3545. Oral dose of mutant strain was 1×10 8 CFU and 1×10 9 CFU, the oral dose of the positive control group was 1×10 5 CFU and 1×10 6 CFU. After the challenge, the mice were fed for 4 weeks, and their growth status was observed and recorded e...
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