Method for producing blattella germanica allergen Bla g 7 protein in bacilliform virus-insect expression system

Abstract

The invention discloses a method for producing German cockroach allergen Bla g 7 protein in a rhabdovirus-insect expression system, comprising the following steps: total RNA is extracted from the tissue of the German cockroach, the total RNA is reversely transcripted into cDNA, a coding gene of German cockroach allergen Bla g 7 is obtained by the method of PCR; the coding gene is cloned to a rhabdovirus carrier, insect cells are transformed for expression; the method of affinity chromatography is adopted for purification, thus obtaining the German cockroach allergen Bla g 7 protein. The protein is different from Bla g 7 protein of foreign sources in terms of 2 amino acid locuses. In the invention, the rhabdovirus-insect expression system is adopted to produce German cockroach allergen Blag 7 protein, thus avoiding influence on Bla g 7 allergen structures in different expression systems (such as bacterial and microzyme); in addition, the invention provides the method of affinity chromatography, thus solving the problem that the Bla g 7 allergen has complex purification process.

Description

A method for producing German cockroach allergen Blag7 protein in baculovirus-insect expression system technical field The invention belongs to the technical field of genetic engineering, and in particular relates to a method for producing Blag7 protein of the German cockroach allergen in a baculovirus-insect expression system. Background technique Allergic diseases are one of the major human diseases. Its incidence rate is currently estimated to account for 30-40% of the world's population, and is increasing at a rate of more than 1% per year, with the most obvious increase in the incidence rate of children. Over the past few decades, the incidence of allergic diseases has increased significantly and rapidly globally, more in Western than in developing countries, and more in urban than in rural areas. Taking allergic asthma (asthma) as an example, there are currently 5.2 million asthmatic patients in the UK: 1.1 million children (1 / 10 of the total population of children)...

AI Technical Summary

Problems solved by technology

The allergen is currently expressed in E. coli and yeast, but there is no report of expression in the baculovirus-insect system

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