An improved protein mmet e 1 for alleviating sensitization to shrimp tropomyosin and its preparation method and application
A technology of tropomyosin and sensitization, which is applied in the field of aquatic food allergy immune regulation, can solve the problems of changing structure and sensory properties, not completely removing allergens, affecting the original value of commodities, etc., and achieving low cost , Relieve food allergic reaction, reduce the effect of allergy to shrimp tropomyosin
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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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