Supercharge Your Innovation With Domain-Expert AI Agents!

Minor polypeptide 102aa and application thereof

A technology of small molecule polypeptide and DNA sequence, applied in small molecule polypeptide 102aa and its carrier and application fields, can solve the problems that cannot be solved in peripheral nerve regeneration cells and molecular events, and achieve the effect of promoting cell differentiation

Active Publication Date: 2017-01-25
NANTONG MINGNUO ELECTRIC TECH CO LTD
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the theory and clinical practice of nerve regeneration have made great progress, especially the emergence and development of microsurgery, but purely microsurgery for repairing nerves cannot resolve the complex cellular and molecular events in peripheral nerve regeneration

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
  • Minor polypeptide 102aa and application thereof
  • Minor polypeptide 102aa and application thereof
  • Minor polypeptide 102aa and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Cloning vector construction of embodiment 1 recombinant peptide

[0022] The full-length Fyn plasmid was purchased from Shanghai Jikai Gene. The upstream primer was 5'-CCGGAATTCGGACCATGATCCAGGCAGAAGAGTGGTACTTTG-3' (including EcoRI restriction site); the downstream primer was 5'-CCGCTCGAGAAACCACAGTTAAGTTAAAACACAAACCATC-3' (containing XhoI restriction site). PCR technology successfully amplified the 306bp mRNA sequence corresponding to 102aa ( figure 1 ), its DNA sequence is shown in SEQ ID NO.2, and the encoded amino acid sequence of 102aa is shown in SEQ ID NO.1. The amplified fragment was digested by ECOR1 and XhoI and ligated with pCMV-HA vector, the ligated product was transferred into competent Escherichia coli DH5α, clones were selected on the Amp+ agar plate, and the recombinant plasmid was extracted by alkaline lysis method, Identification by ECOR1 and XhoI digestion ( figure 2 ).

Embodiment 2

[0023] Expression identification of embodiment 2 recombinant peptide

[0024] Transfect the correctly connected 102aa polypeptide eukaryotic expression vector into 293T cells, collect samples 48 hours later, lyse with RIPA cell lysate, and immunoblot results prove the expression of this polypeptide ( image 3 ). The main process is as follows:

[0025] Transfection was performed using lifectamine 2000 kit. After digesting the cells in the logarithmic growth phase with trypsin, 5 × 10 cells per well 5 The cell density of each was inoculated in a 6-well plate; when the cell confluency reached 60%, the serum-free medium was replaced and equilibrated for 2 hours; then 2 μg of plasmid DNA was dissolved in 250 μL of serum-free medium and mixed; 4 μL of liposomes were mixed In 250 μL base culture; combine the above two solutions, mix well, and incubate at room temperature for 20 minutes; add 500 μL medium containing liposomes and plasmid DNA to each well, mix gently and place at 3...

Embodiment 4

[0026] Example 4 Differentiation of Shwann cells, detection of undifferentiated PC12 cell process extension function

[0027] Prove in vitro the differentiation-promoting and axon-extending effects of recombinant peptides at the cell level: Taking primary Shwann cells and undifferentiated PC12 cells as examples, cAMP 10uM stimulated primary Shwann cells for 72h and NGF 100ng / ml stimulated undifferentiated PC1272h, and Lipofectamin 2000kit was used to The 102aa polypeptide eukaryotic expression vector and the control group were transiently transfected, and the differentiation of Shwann cells and the morphological changes of undifferentiated PC12 cells were observed after 48 hours. The results were as follows Figure 4 and Figure 5 , showing that compared with the empty control group, the 102aa polypeptide eukaryotic expression group significantly promoted the differentiation of Shwann cells and the extension of undifferentiated PC12 cells.

[0028] Footprint experiment of 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

No PUM Login to View More

Abstract

The invention discloses a minor polypeptide 102aa and application thereof. The minor polypeptide 102aa has the amino acid sequence shown as SEQ ID NO.1. A biological engineering technology is used; a section of DNA sequence corresponding to 102 amino polypeptides is subjected to genetic recombination to obtain a pCMV-HA eukaryotic expression vector. Through enzyme digestion and sequence analysis, after the recombination success is proved, the eukaryotic expression recombinant polypeptides transfect nerve cells; the immunoblotting proves the protein expression of the polypeptides; the polypeptide recombination is realized. Then, the peripheral nerve regeneration promoting function of the polypeptides is studied; cytology and rat in vivo experiments show that the polypeptide has the effects of promoting Schwann cell differentiation, axonal regeneration and moving function recovery.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to the small molecule polypeptide 102aa and its carrier and application. Background technique [0002] Peripheral nerve injury has become a common and frequently-occurring disease in clinical practice. It is caused by traffic accidents, diseases or surgery, and often leads to various complications, such as movement disorders, sensory disorders and neuropathic pain, which seriously affect the quality of life of patients. In recent years, the theory and clinical practice of nerve regeneration have made great progress, especially the emergence and development of microsurgery, but purely microsurgery for nerve repair cannot resolve the complex cellular and molecular events in peripheral nerve regeneration. Therefore, it is necessary to have a deeper understanding of all aspects of nerve regeneration in order to improve the level of clinical application. [0003] After perip...

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): C07K14/47C12N15/12C12N15/85A61K38/17A61P25/02
CPCA61K38/00C07K14/47
Inventor 沈爱国刘永华吴玮杰王友华
Owner NANTONG MINGNUO ELECTRIC TECH CO LTD
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com