Nucleotid sequence of human interleukin 21 and method and application of producing matured human interleukin 21

A technology of interleukin and nucleotide sequence, which is applied in the field of pharmaceutical application to achieve the effect of easy separation and purification

Inactive Publication Date: 2006-11-29
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, no prokaryotic expression system of human interleukin 21 has been prepared b

Method used

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  • Nucleotid sequence of human interleukin 21 and method and application of producing matured human interleukin 21
  • Nucleotid sequence of human interleukin 21 and method and application of producing matured human interleukin 21
  • Nucleotid sequence of human interleukin 21 and method and application of producing matured human interleukin 21

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Establishment of Human Interleukin 21 Mature Protein Coding Gene and Its Efficient Prokaryotic Expression System

[0028] The specific operation includes the following steps:

[0029] 1. Activation of human T lymphocytes and extraction of total RNA: Take 2 parts of peripheral venous blood from normal volunteers, separate peripheral blood mononuclear cells (PBMCs) with lymphocyte separation medium, mix and resuspend in a medium containing 10% calf serum RLM-1640 complete medium, add PHA 5μg / 10 6 cells, cultured for 48 hours. Activated cells were collected. Add 1ml of Tritrol to the cells after centrifugation, add 200μl of chloroform after lysis for 2 minutes, vigorously mix, centrifuge at 12000rpm, absorb the water phase and precipitate with isopropanol, finally dissolve in RNase-free water, denature at 95°C and store at -20°C .

[0030] 2. RT-PCR amplification of the human interleukin-21 mature protein gene coding region: perform reverse transcription reaction (42° ...

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Abstract

The invention discloses a human interleukin 21 nucleotide sequence and the method to produce the mature interleukin 21 and the application. The nucleotide sequence is SEQ ID No.1. The manufacture includes the following steps: making SEQ ID No.1, inducing into expression carrier to form reconstruction carrier, transforming the carrier into prokaryotic cell to cultivate and multiple, taking inducing, distilling, purification, the mature human interleukin 21 of SEQ ID No.2 would be gained. The invention could directly express human interleukin 21 in prokaryotic cell. It could be used to produce gene engineer medicine.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and relates to a nucleotide sequence for expressing mature human interleukin-21 in prokaryotic cells, and also relates to a method for producing mature human interleukin-21 using the nucleotide sequence and the Applications of sequences in pharmaceuticals. Background technique [0002] Interleukin 21 (Interleukin 21, IL-21) is a newly discovered cytokine in recent years, which is highly homologous to interleukin 2, interleukin 15 and interleukin 4, and has a wide range of biological activities, mainly reflected in Co-stimulate T lymphocytes and B lymphocytes, activate natural killer cells (NK), enhance the body's specific and non-specific immune functions, thereby exerting the effects of anti-tumor and immune regulation. It is a new type of biological agent with good development prospects. drug. [0003] Interleukin 21 can only be secreted by activated T lymphocytes. The relevant foreign res...

Claims

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Application Information

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IPC IPC(8): C12N15/24C07K14/54C12N15/09C12N15/70C12N1/21A61K38/20A61K48/00A61P35/00A61P37/00
Inventor 陈国兵汤卫国吴梧桐
Owner CHINA PHARM UNIV
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