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Representation method for recombinant american cockroaches allergen protein

An allergen protein and expression method technology, which is applied in the field of recombinant protein expression, can solve the problems of patients' therapeutic allergic reactions, differences in relative potency and specific antigen levels, and difficult standardization and quantification, so as to avoid non-singleness and Difficult to standardize effects

Inactive Publication Date: 2008-07-16
SHANTOU UNIV MEDICAL COLLEGE
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  • Abstract
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AI Technical Summary

Problems solved by technology

Traditional immunotherapy uses cockroach extracts, which contain a variety of antigenic components. There are significant differences in their relative potency and specific antigen levels. It is difficult to standardize and quantify, and it is easy to cause therapeutic allergic reactions in patients, thus limiting the immune response. therapeutic application

Method used

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  • Representation method for recombinant american cockroaches allergen protein
  • Representation method for recombinant american cockroaches allergen protein
  • Representation method for recombinant american cockroaches allergen protein

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Embodiment 1

[0047] 1 Materials and methods

[0048] 1.1 Materials

[0049] 1.1.1 Periplaneta americana

[0050] The adults of Periplaneta americana were collected from residential areas in Shantou City, freshly sampled, frozen in liquid nitrogen, and then stored at -70°C.

[0051] 1.1.2 Strains and vectors

[0052] Escherichia coli JM109, BL21(DE3) strains, and pET-28a expression vectors were preserved by the Allergy Research Institute of Shantou University, and the pMD18-T cloning vector was purchased from TAKARA Bioengineering (Dalian) Co., Ltd.

[0053] 1.1.3 Main enzymes and reagents

[0054] Ex Taq Enzyme (TaKaRa), DNA Rapid Recovery Kit, Plasmid Small Extraction Kit (QIAGEN), Restriction Enzyme (NEB); T 4 DNA Ligase, ImProm-II TM Reverse transcription kit, agarose (Promega); TRIZOL Reagent (GIBCO BRL); peroxidase-labeled goat anti-human IgE (ε) (KPL); liquid DAB+substrate luminescent system (DAKO); His Bind The rest of the conventional reagents in the protein purification kit...

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Abstract

The invention relates to the field of biotechnology and discloses an expression method of a recombinant periplaneta Americana allergen protein. The invention aims at solving the shortcoming of the immune tolerance therapy of cockroach allergen leaching liquor, total RNA is extracted from common cockroach species periplaneta Americana tissues for obtaining the coding genes of major allergen genes of Per a 1.0101, Per a 1.0102, Per a 1.0104 and Per a 7.01 of the periplaneta Americana by amplification, the low-temperature induction expression of the recombinant cockroach allergen protein in escherichia coli by adopting genetic engineering means can be used for the diagnosis and the treatment of hypersensitivity diseases which are caused by cockroaches and can avoid the obstacles of the non-single property and the difficult standardization of natural extracts.

Description

[0001] The present application is a divisional application entitled "Recombinant Periplaneta americana Allergen Protein Expression Method", the application date is July 21, 2006, and the application number is 200610036646.0 technical field [0002] The invention relates to the field of biotechnology, in particular to a method for expressing recombinant proteins. Background technique [0003] 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. In allergic diseases, bronchial asthma (referred to as asthma) accounts for about 1 / 4 of all cases. In 2004, the World Health Organization announced in the "Global Asthma Burden Report" that there are 300 million asthma patients in the world. The incidence of asthma in my country is 0.5% to 1.5%, and there are about...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/70C07K14/435A61K38/17A61P37/08
Inventor 张忠芳何韶衡
Owner SHANTOU UNIV MEDICAL COLLEGE
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