Method for producing recombinant dermatophagoides farinae allergen Der f1 and Der f2 fusion protein

Abstract

The invention discloses a method for producing recombinant dermatophagoides farinae allergen Der f1 and Der f2 fusion protein and relates to the technical field of construction of dermatophagoides farinae allergen gene by a biological technology. The method comprises the following steps of: obtaining a coding gene by extracting the total RNA of the dermatophagoides farinae and adopting PCR (Polymerase Chain Reaction) amplification, cutting glue to recover a product, named as Der fm, cloning to pMD19-T vectors, subcloning to expression vectors pET-28a(+), converting the vectors to escherichia coli, carrying out inducible expression by using IPTG (isopropyl-Beta-d-thiogalactoside). The method disclosed by the invention has the advantages that the consumed time is short, the process is simple, the cost is lower, the components such as a solvent can be thoroughly removed, the purity of the allergen can be increased fundamentally, the occurrence of side reaction in immunological therapy is avoided, the purity is high, the classification is clear, the accuracy of diagnosis can be increased, and a standard product is expected to be provided for novel immunological therapy.

Description

Technical field: [0001] The invention relates to the technical field of constructing dust mite allergens by biological technology, in particular to a method for producing fusion proteins of recombinant dust mite allergens Der f1 and Der f2. Background technique: [0002] There are many kinds of dust mites, which widely exist in the living and working environment of human beings. Their excreta, metabolites and mite bodies all have strong allergenicity. It is estimated that about 10% of the world's population is allergic to dust mites, and about 80% % of exogenous asthma is caused by dust mites. Scholars in my country used the crude extract of dust mite to conduct skin prick test on patients with allergic diseases. 47-92.11% of adult asthmatic patients, 51.64-78.85% of asthmatic children, 80% of asthmatic patients About 70% of patients with rhinitis and about 70% of patients with chronic urticaria were positive. In 1998, the direct and indirect economic losses caused by asthma i...

AI Technical Summary

Problems solved by technology

[0004] Therefore, the quality of allergens is crucial to the diagnosis and treatment of allergic diseases. The allergens used for immunodiagnosis and treatment should be pure products rather than crude infusions. However, dust mite allergens mainly exist in excreted The use of biochemical methods to purify dust mite allergens from food and leather shells is time-consuming, cumbersome, and costly, and it is impossible to completely remove solvents and other components, which cannot fundamentally improve the purity of allergens and avoid immunotherapy. The occurrence of side reactions, with the development and maturity of molecular biology technology, the development of genetic engineering technology to produce recombinant allergens has become the mainstream of allergology research. Its high purity and clear classification can improve the accuracy of diagnosis and is expected to provide Novel immunotherapy provides standardized products

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