NmpC subunit vaccine of salmonella paratyphi A and preparation method thereof

A Salmonella and Paratyphoid technology, applied in the field of Salmonella Paratyphi A subunit vaccine and its preparation, can solve the problems of no preparation of Salmonella Paratyphi A vaccine, few types of NmpC, etc., and achieve easy cultivation and fermentation, and low production cost , the effect of simple structure

Inactive Publication Date: 2011-10-05
NAT INST FOR FOOD & DRUG CONTROL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few known NmpC species, so far, there is no report that NmpC protein is used to prepare Salmonella paratyphi A vaccine

Method used

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  • NmpC subunit vaccine of salmonella paratyphi A and preparation method thereof
  • NmpC subunit vaccine of salmonella paratyphi A and preparation method thereof
  • NmpC subunit vaccine of salmonella paratyphi A and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1: the preparation of Salmonella paratyphi A membrane protein NmpC

[0037] 1. Acquisition of the membrane protein NmpC gene of Salmonella paratyphi A

[0038] Using the genome of Salmonella paratyphi A 50973 strain (source: China Medical Bacteria Collection Management Center) as a template, entrust a DNA synthesis company to synthesize upstream and downstream primers

[0039]NmpC-f: 5'-GGGAATTCCATATGGCCGAGGTATATAACAAAGACG-3';

[0040] NmpC-r: 5'-CCGCTCGAGTTAGTGGTGGTGGTGGTGGTGGAACTGGTAGTTCAGACCA-3'.

[0041] Among them, NmpC-f and NmpC-r are used to amplify the NmpC protein gene, and introduce NdeI and XhoI restriction sites and 6×His tags. PCR amplification was performed using Taq plus enzyme: pre-denaturation at 94°C for 10 min; 30 cycles (94°C for 30 s, 52°C for 30 s, 72°C for 1 min); 72°C for 7 min. Amplified products were identified by 1% agarose gel electrophoresis.

[0042] 2. Construction of recombinant expression vector NmpC-pET30a

[0043] Afte...

Embodiment 2

[0052] Embodiment 2: Animal immunization experiment of NmpC protein

[0053] 1. Animal immunity

[0054] BALB / c mice were divided into 3 groups, 20 in each group, 10 of which were blood collected, and 10 were challenged. Immunization was carried out at 0, 2, and 4 weeks by intramuscular injection. Group A is healthy mice; Group B uses aluminum adjuvant; Group C uses NmpC protein + aluminum adjuvant. The immunization dose is 0.2ml per mouse, containing 10μg of antigen and 0.2mg of aluminum adjuvant. Serum collection was performed 10 days after each immunization. The first and second time blood was collected from the canthus venous plexus, and the third time blood was collected from the eyeball. Serum total IgG antibody was measured by ELISA method, and the inter-group differences of each group of data were analyzed by SPSS software. The results showed that the titer of the mouse IgG antibody against NmpC protein was very high.

[0055] 2. Antivirus protection experiment ...

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Abstract

The invention discloses an NmpC subunit vaccine of salmonella paratyphi A and a preparation method thereof. The vaccine comprises an effective dose of outer membrane proteins NmpC of salmonella paratyphi A and a vaccine adjuvant. Amino acid sequences of the outer membrane proteins NmpC are shown as SEQ ID NO.2. In the preparation method, the vaccine is obtained by obtaining the outer membrane proteins NmpC through in-vitro expression and then mixing the outer membrane proteins NmpC with the vaccine adjuvant. The subunit vaccine provided by the invention has the advantages of low production cost and high yield of target proteins; and the subunit vaccine provided by the invention has important significances for producing NmpC subunit vaccines on a large scale and preventing the infection of the salmonella paratyphi A.

Description

technical field [0001] This article belongs to the field of biotechnology, and relates to a subunit vaccine of Salmonella paratyphi A and a preparation method thereof. Background technique [0002] Typhoid fever is an acute infectious disease, including typhoid fever caused by Salmonella Typhi and paratyphoid fever caused by Salmonella paratyphi A, Band C (C 2005). Common symptoms of patients include: headache, anorexia, abdominal pain, constipation or diarrhea, dry cough, apathetic expression, eosinophil decrease or disappearance, and symptoms such as sore throat, epistaxis, and tongue coating may also occur. Patients who do not receive timely treatment will develop poisoning, and may develop ileal perforation, gastrointestinal bleeding, coma, fainting, pneumonia, nephritis, etc. Due to the widespread existence of clinical drug-resistant strains and multi-drug resistant strains of Salmonella paratyphi A, the outbreak of paratyphi A occurred. Therefore, paratyphoid fever h...

Claims

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Application Information

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IPC IPC(8): C12N15/70C07K14/255A61P31/04C12N15/31C12R1/19A61K39/112
CPCY02A50/30
Inventor 曾明王斌梁昊宇计国欣王恒樑
Owner NAT INST FOR FOOD & DRUG CONTROL
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