Preparation of haliotis divericolor supertexta NF-kappa B antibody and detection method thereof

Abstract

The invention relates to a preparation and detection method for Haliotis diversicolor NF-KappaB(Ab-Rel) antibody. The oligonucleotide primers used for amplifying the RHD and TD fragments are designed according to the Ab-Rel full-length cDNA sequence so as to obtain the encoding frame expression vectors which are pGEX-RHD and pGEX-TD; the expression and identification of the escherichia coli recombinant protein are transformed; the escherichia coli recombinant protein is purified using the gel affinity column before the RHD and TD two polyclonal antibodies of Ab-Rel are obtained by injecting the rabbits and the titer of the antibody is detected using the indirect sandwich ELISA; the RHD and TD of Ab-Rel are subject to the protein Western blotting detection using two antibodies and also the double antibodies and the secondary antibody combination indirect sandwich enzyme-linked immunoreaction assay (ELISA) can be utilized to detect the RHD and TD of Ab-Rel. The invention provides a detection technology for cultured shellfish, in particular for Haliotis diversicolor immune factors.

Description

technical field [0001] The invention belongs to the preparation and detection method of immune factor antibodies of shellfish in invertebrates. Background technique [0002] Nuclear factor-kappa B (Nuclear factor-kappa B, NF-κB) is an important immune factor, which is a family with several members. In the late 20th century, Sen and Baltimore first discovered this factor from the nuclear extract of B lymphocytes. At first, it was considered to be a globulin that can specifically bind to the enhancer of the immunoglobulin κ chain gene and promote the expression of the κ chain gene. ((SenR and Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein NF-κB by aposttranslational mechanism. Cell 1986; 46:705-716.). In the following 20 years, NF-κB and its mediated signaling pathways have been obtained Extensive and in-depth research has been carried out. A large number of studies have found that NF-κB can specifically bind to the κB site of the promoter or enhan...

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Problems solved by technology

The application did not fully disclose the full-length sequence of the Abalone NF-κB gene and the specific sequences of the primers for the two domains of the Abalone NF-κB, so it was difficult to achieve the purpose of preparing the Abalone immune factor antibody

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