Biological inhibitor for silver carp surimi gel degradation
A technology for silver carp and fish meat is applied in the field of biological inhibitors for the deterioration of silver carp surimi gel, can solve the problems of high cost, less research, unfavorable digestion and absorption, etc., and achieves high thermal stability, high acid and alkali resistance, specificity strong effect
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[0032] A biological inhibitor for silver carp surimi gel deterioration, comprising the aforementioned shark-derived single domain antibody. The preparation method of the shark-derived single domain antibody comprises the following steps:
[0033] 1) Immunize the hinged shark with silver carp myofibril-binding serine protease as an antigen, and measure the immune response of the shark; collect peripheral blood lymphocytes of highly immune sharks, extract RNA and synthesize a cDNA library, and use gene recombination technology to construct phage The plasmid vector was introduced into Escherichia coli to construct the shark-derived single domain antibody gene text library and identify the capacity of the text library.
[0034] 2) Use helper phagemids to infect the single-domain antibody gene text library, use silver carp fish myofibril-binding serine protease as the antigen, use phage display technology to pass multiple rounds of panning, and obtain the single-domain antibody wit...
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[0036] Specific example: Preparation of silver carp fish meat myofibril-binding serine protease (MBSP) specific shark-derived single domain antibody and inhibition of silver carp surimi gel degradation effect
[0037] 1. Construction of MBSP shark-derived single-domain antibody text library for silver carp meat
[0038] The silver carp meat MBSP prepared in the laboratory was fully mixed with Freund's adjuvant 1:1, and then subcutaneously injected into the dorsal fin to immunize the mouth shark. The immunization was boosted once every two weeks, and the shark serum was collected after four times of immunization. Response effect of MBSP serum; collected shark peripheral blood lymphocytes that responded efficiently after immunization, extracted total RNA by TRIzol method, identified RNA quality by Experion, synthesized cDNA library using SuperScript system after TURBO TMDnase treatment, designed single-domain antibody bidirectional primers, and passed PCR Amplify the DNA product...
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