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

Fusion protein prepared from dog albumin, dog interferon gamma and dog interferon alpha and preparation method thereof

A technology of fusion protein and canine interferon, applied in interferon, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of high cost of interferon, unfavorable clinical application, small molecular weight of interferon, etc.

Inactive Publication Date: 2018-11-23
WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 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, and at the same time, the cost of polyethylene glycol fusion interferon is very high. High, not conducive to clinical application

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
  • Fusion protein prepared from dog albumin, dog interferon gamma and dog interferon alpha and preparation method thereof
  • Fusion protein prepared from dog albumin, dog interferon gamma and dog interferon alpha and preparation method thereof
  • Fusion protein prepared from dog albumin, dog interferon gamma and dog interferon alpha and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] A fusion protein composed of canine albumin, canine interferon gamma and canine interferon alpha, the preparation method of which is as follows:

[0091] 1. Acquisition and amplification of canine albumin (Alb), canine interferon gamma (IFN-gamma) and canine interferon alpha (IFN-α) target genes

[0092] Primer design:

[0093] Design synthetic primers according to the target gene sequence reported in Genebank, see Table 1, introduce BamHI restriction site and Linker sequence into the upstream primer and downstream primer of canine albumin, respectively, and introduce the upstream primer and downstream primer of canine interferon gamma The Linker sequence was introduced respectively, and the Linker sequence and the XhoI restriction site were respectively introduced into the upstream primer and downstream primer of canine interferon alpha.

[0094] Table 1 PCR amplification primers

[0095]

[0096] RT-PCR to obtain the target gene:

[0097] RNA was extracted from ...

Embodiment 2

[0132] A fusion protein composed of canine albumin, canine interferon gamma and canine interferon alpha, the others are the same as in Example 1, except that the competent cells of Escherichia coli BL21(DE3) are replaced by BL21(DE3) with pGro7 plasmid ) competent cells. The SDS-PAGE electrophoresis result of the fusion protein is compared with that of Example 1, and the dominant expression band at about 126.2KD in the supernatant is relatively thick, indicating that after the introduction of molecular chaperone pGro7, the expression of the target protein in the supernatant is better, and it is 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.

[0133] The BL21(DE3) competent cells carrying the pGro7 plasmid were purchased from Shanghai Inshore Science & Tec...

Embodiment 3

[0135] A fusion protein composed of canine albumin, canine interferon gamma and canine interferon alpha, the preparation method of which is as follows:

[0136] 1. Acquisition and amplification of canine albumin (Alb), canine interferon gamma (IFN-gamma) and canine interferon alpha (IFN-α) target genes

[0137] The canine Alb, canine IFN-γ and canine IFN-α in Example 1 are optimized, and the canine Alb, canine IFN-γ and canine IFN-α target genes are artificially synthesized. After optimization, the nucleotide sequences of the three are respectively as follows SEQUENCE LISTING 400, SEQUENCELISTING 400 and SEQUENCE LISTING 400.

[0138] 1.1 Codon optimization

[0139] 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, ye...

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 fusion protein prepared from dog albumin, dog interferon gamma and dog interferon alpha and a preparation method thereof. The fusion protein is formed by dog albumin, dog interferon gamma and dog interferon alpha through the connection by flexible linkers. A recombinant dog pegylated interferon can be obtained through freeze drying after the mixing of the fusion protein andfreeze drying protection agents. The recombinant dog pegylated interferon can obviously prolong the dog interferon half life; the half life is prolonged by 19 times or more through being compared with that of ordinary dog interferon; the broad-spectrum antiviral effects are achieved; the immune response of the dog per se can be improved.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and specifically relates to a fusion protein composed of canine albumin, canine interferon gamma and canine interferon alpha and a preparation method thereof. Background technique [0002] According to the analysis of the current situation of China's pet industry, the market size of my country's pet industry has reached 105.8 billion yuan by the end of 2014, while China's pet medical industry currently only accounts for about 20%. Compared with the relatively mature US market, the output value of its pet medical The ratio reaches 50%, and there is huge room for development in China's pet medical industry. [0003] Dogs are loyal friends of human beings. With the substantial increase in domestic pet dogs and other dogs, the morbidity and mortality of canine diseases continue to increase, causing both spiritual and economic losses to human beings. The close contact between dogs and h...

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): C07K19/00C12N15/62C12N15/70C12N1/21A61P31/12A61P37/04
CPCC07K14/57A61P31/12A61P37/04C07K14/56C07K2319/31C12N15/62C12N15/70C12N2800/22
Inventor 夏兵兵何志远鲍可兵刘家炉单雪芹凡玉芳王亚男
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