A kind of Periplaneta americana allergen protein ba2 and its expression method

A technology of allergen protein and American cockroach, applied in the field of genetic engineering, can solve problems such as difficulty in standardization and complex composition

Active Publication Date: 2021-04-20
SICHUAN GOODDOCTOR PANXI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the diagnosis and treatment of cockroach allergic diseases, the crude infusion of allergens is widely used, but due to its complex composition, it is difficult to standardize and it is easy to cause other allergic reactions mediated by immunoglobulin (IgE) during treatment, Therefore, the clinical use of allergen infusion is limited

Method used

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  • A kind of Periplaneta americana allergen protein ba2 and its expression method
  • A kind of Periplaneta americana allergen protein ba2 and its expression method
  • A kind of Periplaneta americana allergen protein ba2 and its expression method

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Embodiment 1

[0037] 1 Experimental materials

[0038] 1.1 Experimental samples

[0039] Periplaneta americana was provided by Sichuan Good Doctor Pharmaceutical Group Co., Ltd.

[0040] Thirty SPF grade BALB / c mice, female, 6-8 weeks old, weighing 18-22 g, were provided by the Experimental Animal Center of Sichuan University. They were randomly divided into 3 groups: 5 rats in the PBS control group, 10 rats in the crude extract treatment group of Periplaneta americana, and 10 rats in the BA2 protein treatment group. Each group was housed in a cage.

[0041] 1.2 Experimental strains and plasmids

[0042] The strain used was Escherichia coli E. coli BL21(DE3), and the vector used was pET28-a(+) plasmid, which was purchased from Novagen.

[0043] 1.3 Experimental Instruments

[0044] Centrifuge, shaker, electric heating constant temperature water tank, PCR instrument, electrophoresis tank, gel imaging system, Semi-dry transfer system transfer tank (Bio-Rad), mini vertical electrophoresi...

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Abstract

The invention discloses a Periplaneta americana allergen protein BA2 and an expression method thereof: (1) Periplaneta americana allergen protein is obtained by extracting Periplaneta americana RNA, reverse-transcribing it into cDNA, and performing PCR amplification with the cDNA as a template Encoding gene fragment; (2) Cloning the allergen gene into the prokaryotic expression vector, constructing the recombinant expression plasmid pET28a(+)-BA2, and then transferring it into the host bacteria Escherichia coli BL21, inducing its expression with IPTG, and adopting the affinity chromatography method Purify to obtain recombinant Periplaneta americana allergen protein BA2. Pharmacological experiments show that BA2 protein can significantly increase the IgE content in mouse serum (P<0.05), and can significantly increase IL‑4, IL‑5 and IL in mouse alveolar lavage fluid and mouse splenocyte supernatant The content of ‑13 inflammatory factors and the obvious pathological changes in the lung tissue of mice. The method of the invention can obtain the allergen protein with higher purity, which can provide a reference basis for the follow-up research on the Periplaneta americana allergen and the research on potential allergen substances in other animals.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a Periplaneta americana allergen protein BA2 and an expression method thereof. Background technique [0002] Allergic diseases are listed by the World Health Organization as one of the three major diseases to be prevented and controlled in the 21st century. The occurrence of allergic diseases is extremely widespread, and 1 / 3 of the world's population suffers from allergic diseases. Among allergic diseases, bronchial asthma (abbreviated as asthma) accounts for about 1 / 4 of all cases. Asthma can be triggered by many factors, among which allergens, especially indoor allergens, play a very important role. Indoor allergens mainly include animal dander, dust mites and cockroaches. [0003] Reports that cockroaches can cause asthma first appeared in the 1960s. Periplaneta americana is an important allergen of cockroach allergy-induced asthma. Cockroaches, namely Periplaneta ameri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/70C12N15/12C07K1/22
CPCC07K14/43563
Inventor 耿福能岳碧松沈咏梅
Owner SICHUAN GOODDOCTOR PANXI PHARMA
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