Anti-disease prepn. contg. piginterleukin-4,6 fused gene

Abstract

An anti-disease preparation for animals is prepared through cloning the interleukin-4 and -6 gene of pig, fusing them, configuring the eukaryon expression plasmid VPIl 46 of fusion gene, preparing the chitosan nanoparticles by ion exchange method, dissolving the plasmid in the solution of tripolyphosphate and the deacetyl chitosan solution in acetic acid, and proportional mixing. It can be used to immunize the experimental animal by oral application.

Description

technical field

[0001] 1. Biotechnology 2. Molecular Immunology

[0002] Specifically including the cloning and fusion of porcine interleukin-4 and 6 genes and the construction of eukaryotic expression plasmids, the molecular packaging of Tibetan pig interleukin-4 and 6 fusion gene eukaryotic expression plasmids by chitosan nanoparticles and its Applied techniques for enhancing immune response and immune protection in animals. Background technique

[0003] Enhancing the immune response ability of animals against infection and improving the protection rate of vaccines are the most reliable and simple ways for humans to prevent and control infectious diseases. Because traditional drug treatment and routine vaccination cannot fully guarantee the healthy and efficient development of modern animal husbandry, animal immunosuppression and the decline of vaccine immune response and protection levels have seriously restricted the prevention and treatment of infectious diseases. The...

Examples

Embodiment Construction

[0095] 1. Cloning and sequence analysis of porcine interleukin-4 gene

[0096] Sterile blood was collected from the anterior vena cava of pigs, and the lymphocyte separation technology was used to separate the peripheral blood lymphocytes of pigs for in vitro culture. After culturing for 36 hours under the stimulation of Con A, the total RNA of the cultured lymphocytes was extracted, and the total RNA of Tibetan pigs was extracted. A pair of primers for porcine IL-4 gene cDNA amplification was designed. The primer sequences are as follows:

[0097]Primer 1: CGG GAT CCC TAT TCA TGG GTC TCA CCT C

[0098] Primer 2: CGG GAT CCT CAG CTT CAA CAC TTT

[0099] Using total RNA as a template, add 5 μl of 10×RT-PCR reaction buffer, 25mM MgCl to the 50 μl reaction system 2 10μl, 10mM dNTP5μl, RNase Inhibitor (40units / μl) 1μl, AMV RNase XL (5units / uL) 1μl, AMV-Optimized Taq (5units / uL) 1μl, 5′ and 3′ primers 20uM each, total RNA 1~2μl, water to 50 μl. The RT-PCR cycle parameters are:...

technical field

[0001] 1. Biotechnology 2. Molecular Immunology

[0002] Specifically including the cloning and fusion of porcine interleukin-4 and 6 genes and the construction of eukaryotic expression plasmids, the molecular packaging of Tibetan pig interleukin-4 and 6 fusion gene eukaryotic expression plasmids by chitosan nanoparticles and its Applied techniques for enhancing immune response and immune protection in animals. Background technique

[0003] Enhancing the immune response ability of animals against infection and improving the protection rate of vaccines are the most reliable and simple ways for humans to prevent and control infectious diseases. Because traditional drug treatment and routine vaccination cannot fully guarantee the healthy and efficient development of modern animal husbandry, animal immunosuppression and the decline of vaccine immune response and protection levels have seriously restricted the prevention and treatment of infectious diseases. The...

Examples

Embodiment Construction

[0095] 1. Cloning and sequence analysis of porcine interleukin-4 gene

[0096] Sterile blood was collected from the anterior vena cava of pigs, and the lymphocyte separation technology was used to separate the peripheral blood lymphocytes of pigs for in vitro culture. After culturing for 36 hours under the stimulation of Con A, the total RNA of the cultured lymphocytes was extracted, and the total RNA of Tibetan pigs was extracted. A pair of primers for porcine IL-4 gene cDNA amplification was designed. The primer sequences are as follows:

[0097]Primer 1: CGG GAT CCC TAT TCA TGG GTC TCA CCT C

[0098] Primer 2: CGG GAT CCT CAG CTT CAA CAC TTT

[0099] Using total RNA as a template, add 5 μl of 10×RT-PCR reaction buffer, 25mM MgCl to the 50 μl reaction system 2 10μl, 10mM dNTP5μl, RNase Inhibitor (40units / μl) 1μl, AMV RNase XL (5units / uL) 1μl, AMV-Optimized Taq (5units / uL) 1μl, 5′ and 3′ primers 20uM each, total RNA 1~2μl, water to 50 μl. The RT-PCR cycle parameters are:...

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