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

Ganoderma lucidum immunomodulatory protein mutant and application

A technology of immunoregulatory proteins and mutants, applied in the field of bioengineering, can solve the problems of high cytotoxicity and achieve the effect of simple purification process

Active Publication Date: 2021-04-09
安然芝美(上海)生物技术有限公司
View PDF7 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned deficiencies in the prior art, the present invention proposes a mutant of ganoderma lucidum immunoregulatory protein, which can solve the problem that FIP-glu is more toxic to cells in the application. N31S , FIP-gluM T36N and FIP-gluM N31S / T36N The toxicity to cells is significantly lower than that of Ganoderma lucidum immunomodulatory protein (FIP-glu), which can effectively improve its utilization value

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
  • Ganoderma lucidum immunomodulatory protein mutant and application
  • Ganoderma lucidum immunomodulatory protein mutant and application
  • Ganoderma lucidum immunomodulatory protein mutant and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Obtaining Ganoderma lucidum immunoregulatory protein (FIP-glu)

[0028] 1) Extract the genomic DNA of Chizhi. Ganoderma lucidium 50044 was preserved by the Plant Biotechnology Research Center of Shanghai Jiaotong University. Ganoderma lucidum RNA extraction method refers to Tiangen (TIANGEN) RNA extraction kit.

[0029] 2) Amplify the Ganoderma lucidum immunoregulatory protein gene FIP-glu by PCR, and recover and purify the amplified target gene.

[0030]3) The pPIC9K vector and the amplified FIP-glu gene PCR product were digested with EcoR I and Apa I, and the digested plasmid and target gene were recovered. Use the recovered plasmid and target gene with T 4 After DNA ligase ligation, transfer into competent E.coli DH5α cells. The above-mentioned pPIC9K vector and competent E.coli DH5α cells were preserved by the laboratory of Fudan-Jiaotong University-Nottingham Plant Biotechnology Research and Development Center; the above-mentioned EcoR I enzyme, Apa I enzyme a...

Embodiment 2

[0034] Ganoderma lucidum immunoregulatory protein mutant FIP-gluM N31S the acquisition

[0035] Using the nucleotide sequence shown in SEQ ID NO.7 as a template, and using P1 and P2, and P3 and P4 as primers (sequences shown in SEQ ID NO.9-12) respectively, the PCR amplification reaction was carried out. After the resulting products were mixed, a PCR reaction was performed to obtain the Ganoderma lucidum immunoregulatory protein mutant gene FIP-gluM N31S sequence. Recombinant Ganoderma lucidum immunomodulatory protein mutant FIP-gluM N31S The method of obtaining is the same as in Example 1, and its expression product is identified by SDS-PAGE and Western Blot to determine that the resulting protein is the Ganoderma lucidum immunoregulatory protein mutant FIP-gluM N31S , the identification result is as figure 1 Lanes 2 in A and B are shown.

[0036] Table 1 is used for the primer sequence of this embodiment

[0037]

Embodiment 3

[0039] Ganoderma lucidum immunoregulatory protein mutant FIP-gluM T36N the acquisition

[0040] Using the nucleotide sequence shown in SEQ ID NO.7 as a template, P1 and P5 and P6 and P4 as primers (sequences shown in SEQ ID NO.9, 12, 13, 14) respectively, carry out PCR amplification reaction . After the resulting products were mixed, a PCR reaction was performed to obtain the Ganoderma lucidum immunoregulatory protein mutant gene FIP-gluM T36N sequence. Recombinant Ganoderma lucidum immunoregulatory protein mutant FIP-gluM T36N The method of obtaining is the same as in Example 1, and its expression product is identified by SDS-PAGE and Western Blot to determine that the resulting protein is the Ganoderma lucidum immunoregulatory protein mutant FIP-gluM T36N , the identification result is as figure 1 Lanes 3 in A and B are shown.

[0041] Table 2 is used for the primer sequence of this embodiment

[0042]

[0043]

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

No PUM Login to View More

Abstract

Disclosed is a ganoderma lucidum immunomodulatory protein mutant. The ganoderma lucidum immunomodulatory protein mutant comprises FIP-gluMN31S, FIP-gluMT36N and FIP-gluMN31S / T36N, wherein the amino acid sequence and the nucleotide sequence of the FIP-gluMN31S are shown as SEQ ID NO.1 and SEQ ID NO.4 respectively; the amino acid sequence and the nucleotide sequence of the FIP-gluMT36N are as shown in SEQ ID NO.2 and SEQ ID NO.5 respectively; and the amino acid sequence and the nucleotide sequence of the FIP-gluMN31S / T36N are shown as SEQ ID NO. 3 and SEQ ID NO. 6 respectively. According to the mutant, the problem that the toxicity of FIP-glu in application to cells is relatively high can be solved, the toxicity of the modified mutant to the cells is remarkably reduced compared with that of ganoderma lucidum immunomodulatory protein (FIP-glu), and the utilization value of the mutant can be effectively improved.

Description

[0001] This application is a divisional application of application number [201910792979.3] application date [2019 / 8 / 27] title [Ganoderma lucidum immunoregulatory protein mutant and its application]. technical field [0002] The invention relates to a technique in the field of bioengineering, in particular to a mutant of ganoderma lucidum immunoregulatory protein. Background technique [0003] Fungal immunomodulatory protein (fugal immunomodulatory protein, FIP), which was originally isolated from higher basidiomycetes, is a small molecule protein with immune regulation function. After Kino et al. isolated the first FIP ​​(LZ-8 or FIP-glu) from Ganoderma lucidum (Ganodermaspp.) in 1989, researchers successively discovered more FIPs from various edible and medicinal fungi. They constitute a new protein family - FIP. The proteins of the FIP family have various immunoregulatory effects such as anti-tumor, anti-allergic, and stimulating immune cells to produce various cytokines,...

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): C07K14/375C12N15/31C12N15/81A61K38/16A61P35/00A61P35/02C12R1/84
CPCC07K14/375C12N15/815A61P35/00A61K38/00A61P35/02
Inventor 周选围黎刘定吉毛培文李奇嶂
Owner 安然芝美(上海)生物技术有限公司
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