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Application of escherichia coli succinodehydrogenase iron-sulfur protein SdhB

A technology of Escherichia coli succinic acid and Escherichia coli is applied in the field of medicine to achieve the effect of preventing and treating diseases

Active Publication Date: 2013-09-18
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, vaccines related to Escherichia coli have been developed, but due to the large number of serotypes of Escherichia coli, its effect needs to be further improved

Method used

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  • Application of escherichia coli succinodehydrogenase iron-sulfur protein SdhB
  • Application of escherichia coli succinodehydrogenase iron-sulfur protein SdhB
  • Application of escherichia coli succinodehydrogenase iron-sulfur protein SdhB

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation and purification of Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB

[0032] S1. Amplification of the Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB gene.

[0033] S11. Primer design: According to the whole genome sequence of Escherichia coli published by NCBI, design a pair of primers: the nucleotide sequence of the forward primer F is shown in SEQ ID NO3 5'-CCGGAATTCATGAGACTCGAGTTTTCA-3'; the nucleus of the reverse primer R The nucleotide sequence is shown in SEQ ID NO4: 5'-CCGAAGCTTTTACGCATTACGTTGCAACA-3'. In order to facilitate the construction of cloning and expression vectors, restriction endonuclease sites EcoRI and HindⅢ were respectively introduced into the primers. Primers were synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.

[0034] S12. Gene amplification: extract the whole genome DNA of Escherichia coli, and the extraction of the whole genome DNA adopts the bacterial genome DNA mini-ext...

Embodiment 2

[0041] Example 2: Preparation and purification of Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB antiserum.

[0042] S1. Preparation of Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB antiserum:

[0043] Kunming mice were immunized with the Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB purified in Example 1 to obtain SdhB antiserum. The specific steps are as follows: dissolve the lyophilized powder of Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB in an appropriate amount of ultrapure water, and then mix it with an equal volume of Freund's complete adjuvant (lanolin:paraffin oil=1:4v / v, Bacillus Calmette-Guerin is 30mg / mL) mixed evenly, the protein content in it is about 100ug, and then sonicated for 2min to obtain the SdhB antigen, which is injected intraperitoneally; after 14 days, the iron-sulfur protein of Escherichia coli succinate dehydrogenase SdhB was mixed with an equal volume of Freund's in...

Embodiment 3

[0048] Example 3: Passive immune protection of Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB antiserum.

[0049] Kunming rats about 3 weeks old were used as test subjects, and 40 micrograms of the Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB antiserum prepared in Example 2 were injected intramuscularly, and the control group was injected with the same amount of serum from unimmunized rabbits. One hour later, the drug-resistant Escherichia coli pathogenic strain Y17 (1.5×10 8 cfu / only) for infection. After 72 hours, the survival of Kunming mice in each group was observed, and the relative immune protection rate of immunized Kunming mice was calculated (Table 2). The results showed that the mortality rate of Kunming rats passively immunized with Escherichia coli succinate dehydrogenase iron-sulfur protein SdhB antiserum decreased significantly, and the relative protection rate was 93.75%.

[0050] Table 2 Protective effect of Escherichi...

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Abstract

The invention belongs to the field of medical technology, and specifically relates to an application of escherichia coli succinodehydrogenase iron-sulfur protein SdhB. According to active immunization and passive immunization tests, the escherichia coli succinodehydrogenase iron-sulfur protein SdhB and the corresponding antibody have an obvious immune protection effect on the infection of clinical pathogenic bacteria of escherichia coli. Therefore, the escherichia coli succinodehydrogenase iron-sulfur protein SdhB and the corresponding antibody can be used for preparing a medicine for preventing and treating diseases caused by the infection of clinical pathogenic bacteria of animal escherichia coli. The medicine does not generate drug resistance and can effectively prevent and treat the diseases caused by the infection of clinical pathogenic bacteria of escherichia coli.

Description

technical field [0001] The invention relates to the technical field of medicine, and more specifically relates to the application of the iron-sulfur protein SdhB of Escherichia coli succinate dehydrogenase. Background technique [0002] Escherichia coli ( Escherichiacoli ) is usually called Escherichia coli, distributed in nature, most of them are non-pathogenic, mainly attached to the intestines of humans or animals, and are normal flora, a small number of Escherichia coli are toxic and can cause diseases, including humans and livestock Related diseases of poultry. In recent years, Escherichia coli disease has brought more and more losses to the poultry industry. Its clinical characteristics such as difficult to cure, high mortality rate, and easy recurrence are bothering the majority of farmers and veterinary workers at the grassroots level. For example, Escherichia coli can cause an intestinal infectious disease in piglets called porcine colibacillosis. There are three...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/16A61K39/40A61P31/04
CPCY02A50/30
Inventor 彭宣宪李惠程志学
Owner SUN YAT SEN UNIV
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