Prepn. and application tech. for pig interleukin-4 gene anti-diseases prepns.

A technology of interleukin and gene, applied in gene therapy, anti-infective drugs, allergic diseases, etc., can solve problems such as low efficiency and large DNA dose

Inactive Publication Date: 2005-11-09
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since intramuscular injection mainly induces immune response through Th1 cell

Method used

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  • Prepn. and application tech. for pig interleukin-4 gene anti-diseases prepns.
  • Prepn. and application tech. for pig interleukin-4 gene anti-diseases prepns.
  • Prepn. and application tech. for pig interleukin-4 gene anti-diseases prepns.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0071] 1. Cloning of Tibetan pig IL-4 gene (TPIL-4 gene)

[0072] 10 mL of sterile blood was collected from the ear vein of healthy Tibetan pigs, and the peripheral blood lymphocytes of Tibetan pigs were separated, stimulated by adding 10 ug / ml Con A, and incubated in CO 2 Cultured in an incubator at 37°C for 72 hours, extracted the total RNA of Tibetan pigs in different periods (24, 48, 70 hours), and designed a pair of primers for amplifying the cDNA of the Tibetan pig IL-4 gene. The primer sequences are as follows:

[0073] Primer 1: CGG GAT CCC TAT TCA TGG GTC TCA CCT C

[0074] Primer 2: CGG GAT CCT CAG CTT CAA CAC TTT

[0075] 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, primer 1 and primer 2 each 20uM, total RNA 1-2μl, water to 50 μl. RT-PCR cycle parameters are: 50°C, 30min; 94°C, ...

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Abstract

An anti-infectious immunopotentiator for animals is prepared through cloning the interleukin-4 gene of Tibetan pig, configuring its eukaryon expression plasmid VTPIL-4, preparing chitosan nanoparticles from deacetyl chitosan, and using said chitosan nanoparticles for molecular packaging of plasmid VTPIL-4. It can be used for immunizing experimental animal by intramuscular injection.

Description

technical field [0001] 1. Biotechnology 2. Molecular Immunology [0002] It specifically relates to the preparation and application technology of anti-infection DNA preparation of Tibetan pig interleukin-4 gene packaged with chitosan nanoparticles. Background technique [0003] The traditional methods to prevent and control livestock and poultry disease infection are drug treatment and vaccine immunization. Due to the long-term use of chemical drugs and antibiotics to treat diseases, the "selective pressure" of drugs has led to the emergence, spread and spread of pathogen-resistant strains, resulting in many drug-induced diseases and bacterial resistance to antibiotics. Reduced curative effect or even failure. Moreover, a large number of drugs also cause accumulation in the body, the quality of animal products is reduced, and human consumption also damages health. At present, due to the excessive reliance on antibiotics to control infectious diseases, the problem of bacte...

Claims

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Application Information

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IPC IPC(8): A61K48/00
Inventor 武梅高荣李惠吴凯源付满良杨毅章欢刘狄广程驰李化
Owner SICHUAN UNIV
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