Pneumocystis carinii recombinant protein as well as preparation method and application thereof

A technology of recombinant protein and spore bacteria, which is applied in the medical field, can solve the problems of clinical toxic side effects and increase of clinical drug resistance reports, and achieve the effect of simple detection method, good cost and cost control

Pending Publication Date: 2021-08-10
安徽医科大学第四附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned drugs all have great clinical toxic and side effects, and reports of clinical drug resistance related to them are also increasing.

Method used

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  • Pneumocystis carinii recombinant protein as well as preparation method and application thereof
  • Pneumocystis carinii recombinant protein as well as preparation method and application thereof
  • Pneumocystis carinii recombinant protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] A kind of Pneumocystis carinii recombinant protein His-A12 of the present embodiment 1-85 , whose amino acid sequence is shown in SEQ ID NO: 1; encoding the Pneumocystis carinii recombinant protein His-A12 1-85 The nucleotide sequence of is shown in SEQ ID NO:2.

Embodiment 2

[0082] Pneumocystis carinii recombinant protein His-A12 in a kind of embodiment 1 of the present embodiment 1-85 The preparation method, preparation steps are as follows:

[0083] 1. Establishment of Pneumocystis japonicum animal model and DNA extraction

[0084] (1) Establishment of animal models:

[0085] Take 30 Kunming female rats of clean grade and raise them in open cages for 1 week. After adapting to the environment, they are divided into two cages, A and B. Wherein, 20 mice in cage A were subcutaneously injected with dexamethasone sodium phosphate injection in the groin of mice, each of 0.5 mg was injected once every three days, and administered for 8 weeks in total, during which the drinking water (containing 1g / L Tetracycline (cold boiled water to prevent bacterial infection) and litter, and normal pelleted feed; a total of 10 mice in cage B were subcutaneously injected with phosphate buffer saline (PBS) solution in the groin as a control.

[0086] During the expe...

Embodiment 3

[0177] This example is used to illustrate the recombinant protein His-A12 in the above examples 1-85 The immunization procedure of mice and its immune protection effect.

[0178] 1. Immunization program

[0179] (1) Animal selection and grouping: 45 Kunming female mice were randomly divided into three cages, A, B, and C, with 15 mice in each cage. The mice in the three cages were all exposed to the external environment, and they drank unlimited high-pressure sterilized water during the feeding process, and the mouse bedding and food were replaced regularly during the feeding process.

[0180] (2) Immunization scheme: His-A12 1-85 Protein antigen immunization;

[0181] Dosage: 150μg / piece;

[0182] Group A: immune to His-A12 1-85 protein antigen;

[0183] Group B: immune PBS+adjuvant;

[0184] Group C blank control.

[0185] On day 0, before immunization, the sera of mice in three cages were collected by docking their tails as negative controls. Group A will emulsify c...

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Abstract

The invention relates to the technical field of medical treatment, and provides a Pneumocystis carinii recombinant protein. The amino acid sequence of the protein is as shown in SEQ ID NO: 1. The invention also provides a preparation method of the Pneumocystis carinii recombinant protein. The method comprises the following steps: establishment of a Pneumocystis animal model, DNA extraction, primer design and PCR amplification, purification of a target gene segment, connection, transformation and identification of the target gene segment and a cloning vector, connection, transformation and identification of the target gene segment and an expression vector, inducing expression of the target protein, SDS-PAGE identification of the target protein, and purification of the target protein. Meanwhile, the invention further provides three applications of the Pneumocystis carinii recombinant protein. The protein has the following advantages: the Pneumocystis carinii recombinant protein can be used for early detection of Pneumocystis infection, and can be used for preparing preventive vaccines, or can be used as a drug for relieving clinical symptoms caused by Pneumocystis caramei infection.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a Pneumocystis carinii recombinant protein and its preparation method and application. Background technique [0002] Pneumocystis carinii is an opportunistic fungus that infects humans and a variety of mammals. The infections it causes are mainly seen in patients with acquired immunodeficiency (HIV), long-term use of adrenocortical hormones, cytotoxic drugs, and patients receiving radiotherapy and chemotherapy. Patients with malignant tumors, leukemia, organ transplantation, etc. In recent years, with the increasing number of AIDS patients, the number of Pneumocystis infection is also increasing. Therefore, it is imminent to detect, treat and prevent Pneumocystis carinii. [0003] Because the clinical symptoms of Pneumocystis infection are not typical, traditional early diagnosis methods include: (1) pathogenic microbiological examination: by hexamine silver staining or Giemsa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C07K1/22C12N15/31C12N15/70G01N33/569A61K39/00A61P31/10A61P11/00
CPCC07K14/37C12N15/70G01N33/56961A61K39/0002A61P31/10A61P11/00G01N2333/37
Inventor 沈继录童彤方亚平徐华铭唐浩
Owner 安徽医科大学第四附属医院
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