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

Recombinant ovine pegylated interferon tau, fusion protein for preparing same and preparation method of fusion protein

A technology of fusion protein and interferon, applied in the field of biogenetic engineering, can solve the problems of small molecular weight of interferon, high cost of interferon, unfavorable application, etc., and achieve the effect of avoiding denaturation and renaturation, improving immune response, and increasing half-life

Inactive Publication Date: 2017-11-17
WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common long-acting interferon on the market is represented by polyethylene glycol fusion interferon, which only partially solves the problem of short half-life due to the small molecular weight of interferon at the molecular weight level. At the same time, the cost of polyethylene glycol fusion interferon is very high. High, not conducive to clinical application in livestock

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
  • Recombinant ovine pegylated interferon tau, fusion protein for preparing same and preparation method of fusion protein
  • Recombinant ovine pegylated interferon tau, fusion protein for preparing same and preparation method of fusion protein
  • Recombinant ovine pegylated interferon tau, fusion protein for preparing same and preparation method of fusion protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] A fusion protein composed of goat interferon gamma and goat interferon tau, the preparation method of which is as follows:

[0074] 1. Acquisition and amplification of target genes of sheep interferon gamma (IFN-γ) and sheep interferon tau (IFN-τ)

[0075] Primer design:

[0076] Synthetic primers were designed and synthesized according to the target gene sequence reported in Genebank, as shown in Table 1. The EcoRI restriction site and Linker sequence were respectively introduced into the upstream primer and downstream primer of sheep interferon gamma, and the upstream primer and downstream primer of sheep interferon tau The Linker sequence and the XhoI restriction site were respectively introduced in.

[0077] Table 1PCR amplification primers

[0078]

[0079]

[0080] RT-PCR to obtain the target gene:

[0081] RNA was extracted from sheep liver tissue, and the target genes of IFN-γ and IFN-τ were obtained by reverse transcription. The gene sequences of the t...

Embodiment 2

[0115] A fusion protein composed of sheep interferon gamma and sheep interferon tau, the others are the same as in Example 1, except that the competent cells of Escherichia coli BL21 (DE3) are replaced by competent cells of BL21 (DE3) with pGro7 plasmid . The SDS-PAGE electrophoresis result of the fusion protein is compared with that of Example 1, and the dominant expression band at about 59KD in the supernatant is thicker, indicating that after the introduction of molecular chaperone pGro7, the expression of the target protein in the supernatant is better, and the obtained The amount of fusion protein is higher. Most of the proteins expressed in E. coli exist in inclusion bodies; by introducing molecular chaperones into the expression strains, the co-expressed proteins can be correctly folded to achieve protein soluble expression.

[0116] The BL21(DE3) competent cells carrying the pGro7 plasmid were purchased from Shanghai Inshore Science & Technology Co., Ltd. / Simbano Biot...

Embodiment 3

[0118] A fusion protein composed of goat interferon gamma and goat interferon tau, the preparation method of which is as follows:

[0119] 1. Acquisition and amplification of target genes of sheep interferon gamma (IFN-γ) and sheep interferon tau (IFN-τ)

[0120] The IFN-γ and IFN-τ in Example 1 are optimized, and the IFN-γ and IFN-τ target genes are artificially synthesized. After optimization, the nucleotide sequences of the two are respectively as SEQUENCE LISTING 400 and SEQUENCE LISTING 400 As shown in .

[0121] 1.1 Codon optimization

[0122] There are 64 genetic codes, but most organisms tend to use a subset of these. Those that are most frequently used are called optimal codons, and those that are not frequently used are called rare or low-usage codons. Virtually every organism commonly used for protein expression or production (including E. coli, yeast, mammalian cells, plant cells, and insect cells) exhibits some degree of difference or bias in codon usage. The ...

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
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention discloses recombinant ovine pegylated interferon tau, fusion protein for preparing the same and a preparation method of the fusion protein. The fusion protein is formed by connecting sheep interferon gamma and sheep interferon tau through a flexible linker, and the recombinant sheep pegylated interferon tau can be obtained from the fusion protein and a freeze-drying protective agent through mixing and freeze-drying. The recombinant sheep pegylated interferon tau can significantly prolong the half-life of the sheep interferon, the half-life is prolonged 10 times or higher compared with that of normal sheep interferon, and the recombinant sheep pegylated interferon tau has a broad-spectrum antiviral effect and can improve immune response of sheep.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a recombinant sheep peginterferon τ, a fusion protein for preparing the peginterferon and a preparation method thereof. Background technique [0002] Animal infectious diseases caused by viruses have seriously restricted the healthy development of the breeding industry in various countries and regions. China is the country with the largest sheep stock, slaughter and mutton production in the world. The mutton sheep industry is also one of the important pillar industries of my country's animal husbandry. . With the continuous development of the sheep breeding industry, it is inevitable to face the problem of diseases caused by viruses. Animal diseases not only cause huge economic losses to sheep farmers, but more seriously, some zoonotic infectious diseases are also Potential threat to human health. [0003] At present, the prevention and treatment of sh...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21A61P31/12A61P37/04C12R1/19
CPCA61P31/12A61P37/04C12N15/62C12N15/70C07K14/555C07K14/57C12N2800/22C07K2319/31C07K2319/00
Inventor 赵俊沈咏舟符修乐夏兵兵徐慕珍程正阳徐文俊单雪芹
Owner WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
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