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

Method for producing human epidermal growth factor and lucid ganoderma immunomodulatory protein and application

An immunomodulatory protein, epidermal growth factor technology, applied in the field of genetic engineering, to achieve the effect of strong activity, convenient use and wide application range

Active Publication Date: 2020-06-16
LUDONG UNIVERSITY
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the literature and patents, there is no application of the fusion of human epidermal growth factor and Ganoderma lucidum immunomodulatory protein, and other studies may not have both medical and cosmetic effects

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
  • Method for producing human epidermal growth factor and lucid ganoderma immunomodulatory protein and application
  • Method for producing human epidermal growth factor and lucid ganoderma immunomodulatory protein and application
  • Method for producing human epidermal growth factor and lucid ganoderma immunomodulatory protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Preparation of fusion gene TEF.

[0042] 1) The nucleotide sequence information of the coding genes corresponding to epidermal growth factor EF (GenBank: M26695.1) and LZ-8 (GenBank: M58032.1) obtained from the gene bank of the National Center for Biotechnology Information (NCBI), The DNA analysis software (Gene Runner) converts the nucleotide sequence information of the coding genes of EF and LZ-8 genes into nucleotide sequences containing optimized rice genetic codes, and at the same time, fuses TDS with codon-optimized EF Create a new gene, denoted as TGF; the codon-optimized LZ-8 and TAT were fused to form a new gene, and the sequence information was sent to Nanjing GenScript Biotechnology Company for artificial synthesis. The nucleotide sequence of the fusion gene after codon optimization was changed by 23.5%, and the genetic code was changed by 74.5%.

[0043] 2) Flexible fusion of TGF and LZ-8.

[0044] The total volume of the PCR reaction was 50 μl....

Embodiment 2

[0045] Example 2: The TEF fusion gene is introduced into a transgene vector to construct a plant transgene vector.

[0046] Combine the above-mentioned target fragment TEF with the transgenic vector ubi preserved in the laboratory, through Bam The HI digestion sites were linked, T4 DNA ligase (purchased from NEB Company), 16 °C, ligated for 12 h, transformed into Escherichia coli TP10 competent cells, and the forward and reverse were identified by PCR. Insert the correct TEF-ubi into Agrobacterium ( Agrobacterium tumefaciens) EHA105.

Embodiment 3

[0047] Example 3: Construction of plant transgene vectors for introduction into Agrobacterium.

[0048] The TEF-ubi identified in the correct direction was transformed into Agrobacterium EHA105 by electric shock. The experimental parameters of the electric shock protocol are: 2.5 KV, electrophoretic impulse 25 µF, and resistance 400 ohms.

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

The invention discloses a method for simultaneously producing a human epidermal growth factor and lucid ganoderma immunomodulatory protein by utilizing plant cells. A gene flexible fusion method is used; the human epidermal growth factor and lucid ganoderma immunomodulatory protein gene sequence with artificially optimized codons are subjected to connection using a modified flexible sequence, so that a brand-new fusion gene sequence is created, and then the brand-new fusion gene sequence is connected to a transgenic plant vector, agrobacterium is transformed, agrobacterium EHA 105 TEF-ubi is obtained and preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2018013. By utilizing the strain disclosed by the invention, efficient expression of the TEF fusion gene in plant cells is realized, and the expressed human epidermal growth factor and lucid ganoderma immunomodulatory protein are independent of each other, exist in the cells together, do not influence each other and have respective biological activity. The product produced by the method has the functions of skin repair, whitening, wrinkle resistance and the like, and can be widely applied to skin care products. Meanwhile, the protein produced by the method has a repairing function on epidermal cells, and can be applied to adjuvant therapy of ulcer, scald and the like.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a recombinant TEF fusion protein using a gene gun method to transfer genes and a gene fusion method, a preparation method thereof, and applications in cosmetology and medicine. Background technique [0002] With the development of genetic engineering technology, some small molecular polypeptides with biological activity have been highly valued. Human epidermal growth factor (hEGF), also known as uropregnant, is a small molecule polypeptide composed of 53 amino acids, with a molecular weight of about 6,000 daltons and an isoelectric point of about 4.6. There are three pairs of Disulfide bond, so it is relatively stable to physical and chemical factors such as acid, alkali and heat. hEGF has a wide range of biological effects and can strongly promote the growth of various epidermal tissues. It has been used in medicine for the treatment of burns, ulcers, va...

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): C12N15/62C12N1/21C12N5/10C07K19/00A61K8/64A61K38/18A61K38/16A61Q19/02A61Q19/08A61P17/02C12R1/01
CPCC07K14/485C07K14/37A61K8/64A61K38/1808A61K38/16A61Q19/02A61Q19/08A61P17/02C07K2319/00
Inventor 张秋胜赵燕吕若萱徐丙莲唐清美刘雪
Owner LUDONG UNIVERSITY
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