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Recombinant lactococcus lactis for secreting and expressing Dermatophagoides pteronyssinus allergen Derp2 and application of recombinant lactococcus lactis

A technology for secreting and expressing Lactococcus lactis, which is applied to the recombinant Lactococcus lactis that secretes and expresses the house dust mite allergen Derp2 and its application field, which can solve the problems of low purity, hindering the clinical application of dust mite allergen reagents, and complicated operations

Inactive Publication Date: 2014-03-19
JIANGNAN UNIV
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  • Claims
  • Application Information

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Problems solved by technology

At present, the crude extract of dust mite used clinically for diagnosis and treatment contains a large amount of non-specific impurities, which seriously hinders the clinical application of dust mite allergen reagents
In order to obtain high-quality allergens, isolate and purify allergens from natural products (patent application number: 200410022961.9), the operation is complicated and the purity is not high, and the repeatability is poor; : 201210139240.0), yeast and other expression systems to express the allergen Derp2 is considered a feasible strategy, but its safety and subsequent purification of allergens limit the practical application of these expression systems

Method used

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  • Recombinant lactococcus lactis for secreting and expressing Dermatophagoides pteronyssinus allergen Derp2 and application of recombinant lactococcus lactis
  • Recombinant lactococcus lactis for secreting and expressing Dermatophagoides pteronyssinus allergen Derp2 and application of recombinant lactococcus lactis
  • Recombinant lactococcus lactis for secreting and expressing Dermatophagoides pteronyssinus allergen Derp2 and application of recombinant lactococcus lactis

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Embodiment Construction

[0026] The present invention will be described in more detail below in conjunction with specific embodiments.

[0027] 1. According to the genome data of Lactococcus lactis, analyze the parameters such as codon preference and GC content of Lactococcus lactis, the natural gene sequence of the main allergen Derp2 derived from the house dust mite (Dermatophagoides pteronyssinus) The modified allergen gene The sequence is SEQ ID No.2 (genebank: AF276239.1) for codon optimization, and the gene mderp2 that can be highly expressed in Lactococcus lactis is obtained, and the sequence of the gene is SEQ ID No.1;

[0028] 2. The optimized gene mderp2 was cloned into the expression plasmid pNZ8148 of Lactococcus lactis (gifted by Professor Zhong Jin of the Chinese Academy of Sciences) to obtain the secretory expression plasmid pNZ8148-SD, and the expression plasmid was electrotransformed into the host strain L. lactis NZ9000 ( The recombinant strain L.lactis SD was obtained from the Chine...

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Abstract

The invention relates to an optimized gene mderp2 obtained by carrying out codon optimization onto a gene of primary allergen Derp2 of the Dermatophagoides pteronyssinus, a recombinant expression plasmid pN8148-SD constructed based on the gene for secreting and expressing allergen Derp2 and recombinant lactococcus lactis L.lactis SD (preservation number as CGMCC (China General Microbiological Culture Collection Center) No.8223). Western blot results show that the recombinant lactococcus lactis L.lactis SD disclosed by the invention can realize secretion and expression of the allergen Derp2; meanwhile, an animal experiment result shows that orally-taken recombinant lactococcus lactis L.lactis SD can increase level of regulatory T cells in a mouse spleen, and can lower anaphylactic reaction caused by dust mite allergen Derp2 in the mouse body, and has a better preventive effect.

Description

Background technique [0001] Allergic diseases are clinically common and frequently-occurring diseases, and are currently a major worldwide health problem. Among the many inhaled allergens that cause allergic diseases, dust mites are one of the important factors leading to respiratory allergic diseases. Dust mite is a kind of organism with complex structure and composition. At present, dozens of allergens have been initially identified from hundreds of proteins of dust mite; allergen Derp2 is one of the most important allergens, about 80%- 90% of patients with dust mite allergy develop an allergic reaction to Derp2. Specific immunotherapy (SIT) is considered the most effective treatment for allergies, but successful immunotherapy depends on high-quality allergens. At present, the crude extract of dust mite used clinically for diagnosis and treatment contains a large amount of non-specific impurities, which seriously hinders the clinical application of dust mite allergen reage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N1/21C12N15/74A61K48/00A61K39/35A61P37/08C12R1/01
Inventor 陈卫张秋香艾春青王刚田丰伟刘小鸣赵国忠赵建新张灏郭敏
Owner JIANGNAN UNIV
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