Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Human interleukin-12 recombinant insect virus strain and its prepn

A technology for interleukin and insect virus, which is applied in the field of preparation of recombinant insect virus strains expressing human interleukin-12, and can solve the problems of immune response, the amount of hIL-12, toxicity and the like

Inactive Publication Date: 2002-07-10
WUHAN UNIV
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although studies have shown that hIL-12 has great application prospects and there are many methods for preparing hIL-12, there is a problem with the use of hIL-12. Excessive use will cause the body's immune response and produce toxic effects
Therefore, the use of genetic engineering to obtain hIL-12 for clinical treatment, timely and appropriate provision of patients, is superior to gene therapy, and there is a problem with the safety of introduced viruses in gene therapy
In addition, although some scholars express P35 and P40 subunits separately, the P35 and P40 subunits in hIL-12 are only active through the formation of dimers.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human interleukin-12 recombinant insect virus strain and its prepn
  • Human interleukin-12 recombinant insect virus strain and its prepn
  • Human interleukin-12 recombinant insect virus strain and its prepn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0042] Example 1. Cloning and Identification of P35 Subunit cDNA and P40 Subunit cDNA of Human Interleukin 12

[0043] Collect and culture 20hKB cells stimulated by PDBu (100nM, Sigma), collect the cells, use guanidine isothiocyanate to extract total RNA, synthesize cDNA with MMLV reverse transcriptase, use it as a template and use the following P35 and P40 primers to amplify by PCR respectively. Increased P35 and P40 DNA fragments. P35 Primer: P1 TTGATCA ATG TGT CCA GCG CGC AGCCT

[0044] P2 AGTGACG AGC TAT CTG AAT GCTTCC TAAP40 Primer: P3 AGATCT ATG TGT CAC CAG CAGTTG GT

[0045] P4 TGGATCCCTA AC TGCAGGG CAC AGATG

[0046] The total volume of the PCR reaction was 50 μl. The reaction conditions were: 94°C pre-denaturation for 5 minutes, PCR cycle parameters: 94°C for 60 seconds, 58°C for 90 seconds, 72°C for 120 seconds, 30 cycles, and the last cycle was extended at 72°C for 7 minutes. After 0.8% Agarose electrophoresis detection, a single 667bp P35 cDNA fragmen...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Titeraaaaaaaaaa
Login to View More

Abstract

The present invention discloses a human interleukin-12 recombinant insect virus strain, the recombinant Autographa california multiple nuclear polyhedrosis virus AcNPV-hIL12 virus strain, CCTCC No.2 V200108 and its preparation. The recombinant virus strain is inserted into human interleukin-12 expressing box, and the expressing box includes double promoter sequence, P35 subunit DNA encoding sequence of human interleukin-12 and P40 subunit DNA encoding sequence. P35 subunit gene locates in the downstream of Polyhederin and P40 subunit gene locates in the downstream of P10 promoter. The present invention can express human interleukin-12, and the cell factor is safe and reliable to human body and suitable for treating tumor bacteria, virus and various infections diseases.

Description

technical field [0001] The present invention relates to biotechnology, more specifically relates to human interleukin-12 recombinant insect virus strain (AcNPV-hIL12), and also relates to a preparation method for expressing human interleukin-12 recombinant insect virus strain. Background technique [0002] Human interleukin-12 (human lnterlukin12, hIL-12) is an important cytokine in the human body, also known as natural killer cell stimulating factor (NKSF) or cytotoxic lymphoid maturation factor (CLMF). In 1989, Kobayashi et al. first isolated and purified hIL-12 (Kobayashi M, et al. J Exp Med, 1989ml70:827-845). In 1991, Gubler and Wolf cloned the full sequence of hIL-12 cDNA and expressed it in mammalian cells (Gubler U, et al, Proc Nati Acad Sci USA, 1991, 88:4143-4147; Wolf S F, et al, J Immunol, 1991, 146:3047-3081). hIL-12 is composed of two protein subunits P35 and P40 that are not genetically related to each other through a disulfide bond ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N7/01C12N15/31
Inventor 孟小林徐进平于在林富岩
Owner WUHAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com