Improved protein mMet el for relieving tropomyosin sensitization response of prawns as well as preparation method and application of improved protein

A tropomyosin and sensitization technology, applied in the field of aquatic food allergy immune regulation, can solve the problems of affecting the original value of the commodity, changing the structure and sensory properties, and not being able to completely remove allergens, etc. Allergic reaction, low cost, accurate effect on shrimp tropomyosin allergy

Active Publication Date: 2017-09-01
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the above methods can fundamentally alleviate or control food allergies such as prawns. On the one hand, while eliminating food allergens such as prawns, they cannot completely remove allerge...

Method used

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  • Improved protein mMet el for relieving tropomyosin sensitization response of prawns as well as preparation method and application of improved protein
  • Improved protein mMet el for relieving tropomyosin sensitization response of prawns as well as preparation method and application of improved protein
  • Improved protein mMet el for relieving tropomyosin sensitization response of prawns as well as preparation method and application of improved protein

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Improved protein sequence design

[0025] mMet e 1 improved protein sequence design: based on the amino acid sequence of the new tropomyosin Met e1 of the genus Penaeus, several types of marine fish (Atlantic salmon, grouper, mandarin fish and tuna) that are not allergenic Myosin is the corresponding improved tropomyosin mMet e 1 sequence (SEQ ID NO: 6) obtained based on statistical analysis.

[0026] The comparison of the amino acid sequences of the new Penaeus tropomyosin Met e1, four reference fishes and the improved protein mMet e1 is shown in Table 1.

[0027] Table 1

[0028]

[0029]

[0030]

[0031] 2. cDNA sequence acquisition and synthesis

[0032] The amino acid sequence of the improved tropomyosin was analyzed, and the corresponding cDNA sequence was obtained by reverse transcription and translation, and the corresponding cDNA was synthesized.

[0033] This step was sent to Sangon Bioengineering (Shanghai) Co., Ltd. for completion.

[0034] ...

Embodiment 2

[0048] 1. Improved protein sequence design

[0049] mMet e 1 improved protein sequence design: Based on the amino acid sequence of the new tropomyosin Met e 1 of the genus Penaeus, several types of marine fish (Atlantic salmon, spotted grouper, mandarin fish and tuna) that are not allergic The corresponding improved tropomyosin mMet e 1 sequence (SEQ ID NO: 6) was obtained based on statistical analysis of tropomyosin.

[0050] 2. cDNA sequence acquisition and synthesis

[0051] The amino acid sequence of the improved tropomyosin was analyzed, and the corresponding cDNA sequence was obtained by reverse transcription and translation, and the corresponding cDNA was synthesized.

[0052] 3. Vector construction and protein expression

[0053] Obtain the full-length cDNA gene sequence, and introduce restriction endonuclease EcoRV and HindⅢ enzyme cutting sites at both ends of the sequence, digest and connect with the Pet30(a)+ prokaryotic expression vector, so that the start codon...

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Abstract

The invention discloses an improved protein mMet el for relieving tropomyosin sensitization response of prawns as well as a preparation method and application of the improved protein. The improved protein mMet el has an amino acid sequence shown as SEQ ID NO:1. The preparation method comprises the following steps: (1) contrasting metapenaeus tropomyosin mMet el antigen-specific epitope amino acid sequences and tropomyosin sequences of four marine fishes, performing amino acid sequence analysis, and redesigning antigen epitope amino acid residue sequences so as to obtain the amino acid sequence of the improved protein mMet el; (2) performing reverse transcription and translation so as to obtain cDNA; (3) treating and combining the obtained cDNA and vector plasmids through restriction enzymes so as to obtain a recombinant plasmid vector; (4) introducing the recombinant plasmid vector into host bacteria, and expressing the improved protein mMet el; (5) purifying, thereby obtaining a standard substance of the improved protein mMet el. The improved protein disclosed by the invention has the characteristics of high yield, stable production and low cost.

Description

technical field [0001] The present invention relates to the field of immune regulation of aquatic food allergy, and introduces a new tropomyosin (Met e 1) of the genus prawn (Metapenaeus) as a prototype, through targeted comparison, designed and prepared to relieve prawn The improved protein mMet e 1 preparation method for myosin sensitization aims to provide support for the control of typical aquatic food allergy. Background technique [0002] Food allergy is a typical allergic reaction and an immediate immune response mediated by IgE, which has become a focus of attention in recent years. Shrimp and its products are delicious and nutritious, and are deeply loved by people. However, because of their high allergenicity, they affect the consumption of allergic people. According to statistics, allergic reactions to shrimp and other crustaceans are widely present in various countries around the world, accounting for more than 30% of the total population with food allergies, an...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/70A23L33/18A61K38/17A61P37/08
CPCA23L33/18A23V2002/00A61K38/00C07K14/43509C12N15/70A23V2250/542A23V2200/304
Inventor 傅玲琳王彦波饶志恒周瑾茹
Owner ZHEJIANG GONGSHANG UNIVERSITY
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