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.

Inactive Publication Date: 2019-06-21
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, it is not yet clear whether other methods can be used to attenuate S. choleraesuis

Method used

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  • Attenuated Salmonella choleraesuis s.choleraesuis S1210 and its preparation method and application
  • Attenuated Salmonella choleraesuis s.choleraesuis S1210 and its preparation method and application
  • Attenuated Salmonella choleraesuis s.choleraesuis S1210 and its preparation method and application

Examples

Experimental program
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Effect test

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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Abstract

The invention provides a strain of attenuated salmonella choleraesuis (S. Choleraesuis S1210) which is characterized in that the attenuated salmonella choleraesuis (S. Choleraesuis S1210) is lack of a wbaD gene, is classified and named as S. Choleraesuis S1210, and is preserved in the China center for type culture collection, wherein the preservation number is CCTCC M 2015632, and the preservation time is October 22, 2015. The invention further provides a method for preparing the attenuated salmonella choleraesuis (S. Choleraesuis S1210) and application of the attenuated salmonella choleraesuis (S. Choleraesuis S1210) to vaccine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to attenuated Salmonella choleraesuis S. Choleraesuis S1210 and its preparation method and application. Background technique [0002] With the continuous prevalence of new and old infectious diseases, it is extremely urgent to research and develop effective methods for preventing and treating infectious diseases. In the past two hundred years, vaccines have played a huge role in preventing bacterial or viral infectious diseases. Since vaccines have the advantage of low cost compared with most drugs, continuous research and development of vaccines is very necessary. [0003] In the field of vaccine research, the use of Salmonella as a vector has been extensively studied. However, at present, the virulence genes of Salmonella, especially certain Salmonella (such as Salmonella choleraesuis), have not been fully studied. Over the past two decades, there have been few attenuat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/74A61K39/112A61P31/04C12R1/42
CPCA61K39/0275A61K2039/522C07K14/255C12N15/74
Inventor 孔庆科刘田刘青赵新新
Owner SICHUAN AGRI UNIV
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