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Method for producing American cockroach allergen protein Pera 9 in baculovirus-insect expression system

An allergen protein, Periplaneta americana technology, applied in the field of genetic engineering, can solve the problems of low specific IgE binding activity and complex purification process of natural allergen Pera9 protein

Inactive Publication Date: 2011-06-08
JIANGSU PROVINCE HOSPITAL
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  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to propose a kind of method that produces Periplaneta americana allergen protein in the baculovirus-insect expression system at the shortcoming that above prior art exists, solves the complex and specificity of natural allergen Per a 9 protein purification process The problem of low IgE binding activity

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  • Method for producing American cockroach allergen protein Pera 9 in baculovirus-insect expression system
  • Method for producing American cockroach allergen protein Pera 9 in baculovirus-insect expression system
  • Method for producing American cockroach allergen protein Pera 9 in baculovirus-insect expression system

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Embodiment 1

[0020] 1. The steps of this embodiment are as follows:

[0021] (1) Total RNA was extracted from Periplaneta americana tissue; artificially raised and frozen Periplaneta americana (from Jiangsu Provincial Center for Disease Control) was taken, ground with liquid nitrogen, and Trizol reagent was added at 100 mg tissue / mL Trizol reagent. Let stand for 10 minutes, then add chloroform at 300ul / mL, mix well, let stand for 10min, centrifuge at 12000g for 10min, discard the supernatant, and add isopropanol at 500ul / mL. Mix well, let stand for 10 minutes, centrifuge at 12000g for 10 minutes, discard the supernatant, and the sediment at the bottom of the tube is the total RNA of Periplaneta americana. Add 75% ethanol to suspend the precipitate, centrifuge at 12000 g for 10 min, and discard the supernatant. Dry at room temperature for 5 min, add ribonuclease-free double distilled water to dissolve.

[0022] (2) Using the extracted total RNA of Periplaneta americana as a template, the ...

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Abstract

The invention relates to a method for producing American cockroach allergen protein Pera 9, in particular to a method of producing the American cockroach allergen protein Pera 9 in a baculovirus-insect expression system, and belongs to the technical field of gene engineering. The method produces the American cockroach allergen Pera 9 protein by the baculovirus-insect expression system so as to avoid the problem that a Pera 9 allergen structure is influenced in different expression systems (such as bacteria and yeast). The method provides an affinity chromatography method to solve the problem that the purification process of natural Pera 9 allergen is complicated. The activity of the Pera 9 protein obtained by the baculovirus-insect expression system is as 2.1 times as that of Pera 9 protein obtained by Escherichia coli.

Description

technical field [0001] The invention relates to a method for producing Periplaneta americana allergen protein Per a9, in particular to a method for producing Periplaneta americana allergen protein Per a9 in a baculovirus-insect expression system, and belongs to the technical field of genetic engineering. Background technique [0002] 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. my country's large-scale national surveys in recent years have shown that the incidence of asthma in children has increased significantly compared with 10 years ago, and there are as many as 10 million children with asthma, among which allergies are the main cause (National Children's Asthma Prevention and Control Cooperative Group 2003). In addition, unlike cancer and cardiovascular...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/866C12P21/02
Inventor 魏继福何韶衡
Owner JIANGSU PROVINCE HOSPITAL
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