Prepn. and application of pig interleukin-6 gene anti-infection immunopotentiator

Abstract

An anti-infectious immunopotentiator for animals is prepared through cloning the interleukin-6 gene of Chenghua pig, configuring its eukaryon expression plasmid VPIL-6, preparing chitosan nanoparticles from deacetyl chitosan and using said chitosan nanoparticles for molecular packaging of plasmid VPIL-6. It can be used to immunize the experimental animal by intramuscular injection or oral taking.

Description

technical field

[0001] 1. Biotechnology 2. Molecular Immunology

[0002] It specifically relates to the preparation and application technology of chitosan nano particles packaged into Chinese pig interleukin-6 gene anti-infection immune enhancer. Background technique

[0003] The development of modern animal husbandry is becoming increasingly intensive and large-scale. Due to the wide application of antibiotics in treatment and feed additives, pathogens continue to mutate under the pressure of drugs and immunity (viruses and bacteria are particularly prominent), antigens continue to drift, and drug resistance is increasing. Enhanced; in addition to environmental pollution, intensive feeding and management often put animals in a state of stress, which reduces the immune defense ability of animals, suppresses or weakens the immune response to inactivated vaccination, and reduces the immune protection rate, resulting in infectious diseases in intensive animals. More likely to ...

Examples

Embodiment Construction

[0071] 1. Cloning of Chenghua pig IL-6 gene (PIL-6 gene)

[0072] Take 10mL of blood from the anterior vena cava of healthy Chenghua pigs, separate the peripheral blood lymphocytes of Tibetan pigs, add 10ug / ml ConA to stimulate them, culture them in a CO2 incubator at 37°C for 72 hours, and extract the total RNA of Tibetan pigs in different periods (24, 48, 70 hours). , to design a pair of cDNA amplification primers for Chenghua pig IL-6 gene. The primer sequences are as follows:

[0073] Primer 1: CGG GAT CCA CCA GGA ACG AAA GAG AG

[0074] Primer 2: CGG GAT CCA GGT TTC TGA CCA GAG GAG

[0075] Using total RNA as a template, add 24, 5ul of ddH to the 50μl reaction system 2 O, 4ul 10mM dNTP, 1ul RNase Inhibitor (40units / ul) 2.5ul DTT solution, IL-6 upstream and downstream primers 1ul (15pmol / L), porcine lymphocyte total RNA 1ul, 10ul 5*RT-PCR Buffer, 1ul RT -PCR enzyme mix. RT-PCR cycle parameters are: 50°C, 30min; 94°C, 2min, then perform 30 PCR cycles (denaturation at 9...

technical field

[0001] 1. Biotechnology 2. Molecular Immunology

[0002] It specifically relates to the preparation and application technology of chitosan nano particles packaged into Chinese pig interleukin-6 gene anti-infection immune enhancer. Background technique

[0003] The development of modern animal husbandry is becoming increasingly intensive and large-scale. Due to the wide application of antibiotics in treatment and feed additives, pathogens continue to mutate under the pressure of drugs and immunity (viruses and bacteria are particularly prominent), antigens continue to drift, and drug resistance is increasing. Enhanced; in addition to environmental pollution, intensive feeding and management often put animals in a state of stress, which reduces the immune defense ability of animals, suppresses or weakens the immune response to inactivated vaccination, and reduces the immune protection rate, resulting in infectious diseases in intensive animals. More likely to ...

Examples

Embodiment Construction

[0071] 1. Cloning of Chenghua pig IL-6 gene (PIL-6 gene)

[0072] Take 10mL of blood from the anterior vena cava of healthy Chenghua pigs, separate the peripheral blood lymphocytes of Tibetan pigs, add 10ug / ml ConA to stimulate them, culture them in a CO2 incubator at 37°C for 72 hours, and extract the total RNA of Tibetan pigs in different periods (24, 48, 70 hours). , to design a pair of cDNA amplification primers for Chenghua pig IL-6 gene. The primer sequences are as follows:

[0073] Primer 1: CGG GAT CCA CCA GGA ACG AAA GAG AG

[0074] Primer 2: CGG GAT CCA GGT TTC TGA CCA GAG GAG

[0075] Using total RNA as a template, add 24, 5ul of ddH to the 50μl reaction system 2 O, 4ul 10mM dNTP, 1ul RNase Inhibitor (40units / ul) 2.5ul DTT solution, IL-6 upstream and downstream primers 1ul (15pmol / L), porcine lymphocyte total RNA 1ul, 10ul 5*RT-PCR Buffer, 1ul RT -PCR enzyme mix. RT-PCR cycle parameters are: 50°C, 30min; 94°C, 2min, then perform 30 PCR cycles (denaturation at 9...

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