Multi-species universal ELISA kit for differential diagnosis of foot and mouth disease virus infection
A foot-and-mouth disease virus, differential diagnosis technology, applied in the field of multi-species universal ELISA kits, kits, can solve problems such as unproven domains
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Embodiment 1
[0077] Embodiment 1 recombinant protein [AGL] n Gene design and concatenation
[0078] 1. Materials and methods
[0079] 1.1 Materials
[0080] Recombinant plasmid pET-30a-8BF, expression vector pET-30a(+), Escherichia coli competent cells TG1 and Rosetta(DE3)PlysS were all preserved by our laboratory; cloning vector pJWF was purchased from Beijing Liuhe Huada Gene Technology Co., Ltd., The cloning vector pMD18-T was purchased from Treasure Bioengineering (Dalian) Co., Ltd.; the target protein expression strain BL21-pET-30a-8BF and the empty vector control strain BL21-pET-30a were prepared and preserved by our laboratory. Restriction enzymes BamHI, XhoI, BglII and T4DNA polynucleic acid kinase (T4PNK) were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; T4DNA ligase was purchased from NewEnglandBioLabs (NEB).
[0081] 1.2 Experimental method
[0082] 1.2.1 Design of oligonucleotide chains for the synthesis of B-SPA, B-SPG and B-PPL genes
[0083] The five immun...
Embodiment 2
[0163] Embodiment 2 recombinant protein [AGL] n Prokaryotic expression and identification of
[0164] 1. Materials and methods
[0165] 1.1 Materials
[0166] Ampicillin (Amp) and Kanamycin (Kan) were purchased from Sigma; GelExtractionMiniKit and nitrocellulose membrane (NC membrane) were purchased from Shanghai Huashun Bioengineering Co., Ltd.; protein purification kit Ni-NTAHisHindPurificationSystem kit was purchased from Qiagen; EasySeeWesternBlotKit and WesternMarker were purchased from Beijing Quanshijin Biotechnology Co., Ltd.
[0167] 1.2 Method
[0168] 1.2.1B-SPPAGL gene tandem vector pET-30a(+)-B-SPP[AGL] n build
[0169] For the B-SPPAGL gene on pET-30a(+)-B-SPPAGL, the nucleotide sequences of XhoI, BglII and BamHI enzyme cleavage sites that are convenient for tandem genes are added at the beginning and the end of the design, wherein BglII and BamHI are mutually Homologous enzyme, the B-SPPAGL fragment with cohesive ends recovered after double digestion of pE...
Embodiment 3
[0203] Example 3 Analysis and application of enzyme-labeled protein binding activity to various animal Ig
[0204] 1. Materials and methods
[0205] 1.1 Materials
[0206] 3, 3, 5, 5-tetramethylbenzidine (TMB), rabbit anti-bovine IgG horseradish peroxidase (HRP)-labeled antibody and goat anti-rabbit IgG horseradish peroxidase (HRP)-labeled antibody were purchased from Sigma Company; Brucella antibody rapid detection test kit and Brucella rapid detection test strips were purchased from South Korea Anjie Company; WesternMarker was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; 3ABC antibody detection ELISA kit was purchased from China Agriculture Lanzhou Veterinary Research Institute of the Academy of Sciences; HumanIgM and HumanIgA were purchased from Beijing Bosheng Jingwei Technology Co., Ltd.; goat IgG, pig IgG, guinea pig IgG, human IgG, rabbit IgG, and mouse IgG were purchased from Wanhuashi (Beijing) Biotechnology Co., Ltd.
[0207] 1.2 Method
[0208] 1.2....
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