Allergen protein BA2 of Periplaneta americana and expression method thereof

A technology of allergen protein and Periplaneta americana, applied in the field of genetic engineering, can solve problems such as difficult standardization and complex components

Active Publication Date: 2018-03-06
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 sta

Method used

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  • Allergen protein BA2 of Periplaneta americana and expression method thereof
  • Allergen protein BA2 of Periplaneta americana and expression method thereof
  • Allergen protein BA2 of Periplaneta americana and expression method thereof

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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 an allergen protein BA2 of Periplaneta americana and an expression method thereof. The expression method comprises the following steps: (1) extracting the RNAs of Periplaneta americana, subjecting the RNAs to inverse transcription to obtain cDNAs and carrying out PCR amplification with the cDNAs as a template so as to obtain a coding gene fragment of the allergen protein ofPeriplaneta americana; and (2) cloning an allergen gene into a prokaryotic expression vector to construct recombinant expression plasmid pET28a(+)-BA2, transferring the recombinant expression plasmidinto the host bacterium Escherichia coli BL21, inducing the expression of the recombinant expression plasmid with IPTG and carrying out purifying by using affinity chromatography so as to obtain therecombinant allergen protein BA2 of Periplaneta americana. The results of pharmacological experiments show that the allergen protein BA2 can obviously increase the IgE content (wherein P is less than0.05) of mouse serum, substantially increase the contents of IL-4, IL-5 and IL-13 inflammatory factors in mouse bronchoalveolar lavage fluid and mouse splenocyte supernatant, and result in obvious pathological changes of mouse lung tissue. The method provided by the invention can prepare the allergen protein with high purity and provides referential bases for subsequent research on the allergens of Periplaneta americana and 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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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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