Application process of pig interleukin 2 and 6 gene expression plasmid anti-infectious preparation

Abstract

A process for preparing the anti-infections medicine of pig includes such steps as modifying interleukin 2 gene, configuring its eucaryotic expression plasmid VRIL2S, configuring the recombinant eucaryotic plasmid VRIL6C from pig's interleukin-6 gene and oligonucleotide CpG, preparing chitosan nanoparticles, and using said chitosan nanoparticles for the molecular packing of VRIL2S+VRIL6C plasmid combination. It can be used to immunize pig by injection.

Description

technical field

[0001] 1. Biotechnology 2. Molecular Immunology Background technique

[0002] Interleukin-2 (Interleuki-2, IL-2) is a lymphokine secreted by helper T cells, which plays a key role in the regulation of animal cells and humoral immunity. (1) It can stimulate the proliferation of T lymphocytes and maintain The complete immune system matures and activates key immune molecules of T and B cells, enhancing the killing activity of cytotoxic T cells; (2) IL-2 also induces natural killer (NK) cells and lymphokine activated killer cells ( LAK) growth, involved in the process of destroying tumors. IL-2 can also enhance the activity of natural killer cells (NK cells). Resting NK cells only express the β chain and γ chain of IL-2R, have low affinity for IL-2, and can only respond to high concentrations of IL-2 . Once NK cells are activated, they express the α chain of IL-2R and become high-affinity receptors; the LAK activity induced by high-dose IL-2 mainly comes from ...

Examples

Embodiment Construction

[0057] 1. DNA shuffling of IL-2 gene

[0058] The cloned pig, yak, mouse and human IL2 genes were subjected to DNA enzyme digestion, and 50-100bp nucleotide fragments were separated and recovered; the IL2 gene was re-amplified by two-step PCR without primers and with primers, separated and purified to obtain the shuffled IL2 gene; fragments after shuffling PCR were subjected to 2.0% agarose gel electrophoresis, and 500 bp shuffling fragments were recovered by the agarose gel small molecule DNA extraction kit. Then this DNA fragment was digested with BamHI / EcoRI, and dephosphorylated, BamHI / EcoRI double-digested pGEX 4T-1 vector was digested at T 4 Under the action of DNA ligase, place in a water bath at 16°C overnight. The ligation product was transformed into E.coli DH5α competent cells, the recombinant plasmid was constructed, and the positive cloned gene was identified by enzyme digestion.

[0059] The results are shown in Figures 1 and 2 of the description. Figure 1 sho...

technical field

[0001] 1. Biotechnology 2. Molecular Immunology Background technique

[0002] Interleukin-2 (Interleuki-2, IL-2) is a lymphokine secreted by helper T cells, which plays a key role in the regulation of animal cells and humoral immunity. (1) It can stimulate the proliferation of T lymphocytes and maintain The complete immune system matures and activates key immune molecules of T and B cells, enhancing the killing activity of cytotoxic T cells; (2) IL-2 also induces natural killer (NK) cells and lymphokine activated killer cells ( LAK) growth, involved in the process of destroying tumors. IL-2 can also enhance the activity of natural killer cells (NK cells). Resting NK cells only express the β chain and γ chain of IL-2R, have low affinity for IL-2, and can only respond to high concentrations of IL-2 . Once NK cells are activated, they express the α chain of IL-2R and become high-affinity receptors; the LAK activity induced by high-dose IL-2 mainly comes from ...

Examples

Embodiment Construction

[0057] 1. DNA shuffling of IL-2 gene

[0058] The cloned pig, yak, mouse and human IL2 genes were subjected to DNA enzyme digestion, and 50-100bp nucleotide fragments were separated and recovered; the IL2 gene was re-amplified by two-step PCR without primers and with primers, separated and purified to obtain the shuffled IL2 gene; fragments after shuffling PCR were subjected to 2.0% agarose gel electrophoresis, and 500 bp shuffling fragments were recovered by the agarose gel small molecule DNA extraction kit. Then this DNA fragment was digested with BamHI / EcoRI, and dephosphorylated, BamHI / EcoRI double-digested pGEX 4T-1 vector was digested at T 4 Under the action of DNA ligase, place in a water bath at 16°C overnight. The ligation product was transformed into E.coli DH5α competent cells, the recombinant plasmid was constructed, and the positive cloned gene was identified by enzyme digestion.

[0059] The results are shown in Figures 1 and 2 of the description. Figure 1 sho...

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