Animal genetic engineering interferon alpha and gamma composite preparations and production method and clinical application thereof
A technology of interferon α and genetic engineering, applied in genetic engineering, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of product loss, denaturation and renaturation, and increased cost of enzyme digestion, etc., to increase expression , strong antiviral activity, and strong immunomodulatory activity
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Embodiment 1
[0034] The production method of chicken genetic engineering interferon α and γ compound preparation is specifically carried out by the following steps:
[0035] A. Extract chicken peripheral blood or splenic lymphocytes, culture in vitro and induce with ConA inducer, extract cell total RNA by Trizol method, design specific primers, clone chicken interferon α and γ genes by RT-PCR technology, and use T-A strategy Ligate it to the pGEM-T vector;
[0036] B. Redesign the primers, add restriction sites at the 5′ ends of the upstream and downstream primers, add BamH I and HindIII to chicken interferon α, add Eco I and Sph I to chicken interferon γ, and use T vector as a template to PCR amplify chicken interferon The expression fragments of α and γ genes were respectively connected to the vector pFastBac by double enzyme digestion TM On Dual, the gene expression fragment encoding chicken interferon α is placed under the control of the promoter Polyhedrin promoter, the gene express...
Embodiment 2
[0043] The production method of the compound preparation of porcine genetically engineered interferon α and γ specifically consists of the following steps:
[0044] A. Extract porcine peripheral blood or splenic lymphocytes, culture in vitro and induce with ConA inducer, extract total RNA from cells by Trizol method, design specific primers, clone porcine interferon α and γ genes by RT-PCR technology, and use T-A strategy Ligate it to the pGEM-T vector;
[0045] B. Redesign the primers, add restriction sites to the 5′ ends of the upstream and downstream primers, add EcoR I and HindIII to porcine interferon α, add Nco I and Sph I to porcine interferon γ, and use T vector as a template to PCR amplify porcine interferon The expression fragments of α and γ genes were respectively connected to the vector pFastBac by double enzyme digestion TM On Dual, the gene expression fragment encoding porcine interferon α is placed under the control of the promoter Polyhedrin promoter, the ge...
Embodiment 3
[0052] The production method of the compound preparation of bovine genetically engineered interferon α and γ specifically consists of the following steps:
[0053] A, extract bovine peripheral blood lymphocytes, through in vitro culture and induction of ConA inducer, Trizol method extracts total RNA of cells, designs specific primers, clones bovine interferon α and γ genes by RT-PCR technology, and cloning them by T-A strategy Ligated to pGEM-T vector;
[0054] B. Redesign the primers, add restriction sites to the 5′ ends of the upstream and downstream primers, add BamH I and HindIII to bovine interferon α, add Nco I and Sph I to bovine interferon γ, and use T vector as a template to PCR amplify bovine interferon The expression fragments of α and γ genes were respectively connected to the vector pFastBac by double enzyme digestion TM On Dual, the gene expression fragment encoding bovine interferon α is placed under the control of the promoter Polyhedrin promoter, the gene ex...
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