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

Preparation method of serial antibacterial peptide Alloferon-1 and application of rAlloferon-1-K

A technology of antibacterial peptides and tandems, which is applied in the preparation of tandem antibacterial peptide Alloferon-1 and the application field of rAlloferon-1-K, can solve the problems of low natural expression yield and difficulty in meeting industrialization requirements, and is conducive to popularization and application , the effect of reducing the preparation cost

Inactive Publication Date: 2011-11-16
严杰
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its low natural expression yield is difficult to meet the requirements of industrialization

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
  • Preparation method of serial antibacterial peptide Alloferon-1 and application of rAlloferon-1-K
  • Preparation method of serial antibacterial peptide Alloferon-1 and application of rAlloferon-1-K
  • Preparation method of serial antibacterial peptide Alloferon-1 and application of rAlloferon-1-K

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0024] The preparation method of the tandem antimicrobial peptide Alloferon-1 is:

[0025] 1) Obtain the amino acid sequence of natural Alloferon-1 from GenBank: N-His Gly Val Ser Gly His Gly Gln His Gly Val His Gly-C (Alloferon-1 sequence does not contain arginine and lysine);

[0026] 2) Design a pair of non-mirror-symmetrical single-stranded DNA with complementary cohesive ends; according to the reported amino acid sequence of Alloferon-1 and the codon preference of E. coli, while considering the isolation of Alloferon-1 expressed in tandem, the positive strand sequence: 5'-phosphorylation -CATGGCGTGAGCGGCCATGGCCAGCATGGCGTGCATGGCAAA-3', a trypsin cleavage site lysine codon is set at the 3' end of the positive strand DNA Alloferon-1 gene, the negative strand sequence: 5'-phosphorylation-GCCATGGCCGCTCACGCCATGTTTGCCATGCACGCCATGCTG-3'. Negative-strand DNA interleaved with positive-strand; entrust Invitrogen to synthesize.

[0027] 3) Formation of double strands of the target p...

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 relates to a preparation method and application of an antibacterial peptide. The invention aims to provide a preparation method of an antibacterial peptide Alloferon-1, which has simple steps and high expression efficiency and keeps the in-vitro anti-tumor activity of the antibacterial peptide, and provide application of rAlloferon-1-K. The technical scheme is as follows: the preparation method of the serial antibacterial peptide Alloferon-1 comprises the following steps of: (1) designing a pair of terminal single-stranded DNAs (deoxyribonucleic acids); (2) generating a concatemer in a PCR (polymerase chain reaction) system; (3) carrying out electrophoretic screening on a 14 Alloferon-1 concatemer; (4) constructing a prokaryotic recombinant expression system E.coliBL21DE3pET42a-14Alloferon-1-K; (5) expressing and purifying recombinant protein 14rAlloferon-1-K; and (6) performing trypsin digestion on the 14rAlloferon-1-K, and purifying to obtain the rAlloferon-1-K.

Description

technical field [0001] The present invention relates to the establishment of a peptide tandem prokaryotic expression method, enzymatic cleavage, purification and application of recombinant tandem peptides, in particular to the tandem prokaryotic expression method of the antibacterial peptide Alloferon-1, and the recombinant peptide monomer rAlloferon-1 is obtained by enzymatic digestion with trypsin. 1-K, the antitumor activity of rAlloferon-1-K was tested. Background technique [0002] Antimicrobial peptides are non-antigenic short peptides that widely exist in a variety of organisms. They have broad-spectrum and high-efficiency inhibitory or killing activities against various microorganisms and protozoa. Some antimicrobial peptides also have anti-tumor effects and do not damage normal cells. Therefore, antimicrobial peptides are currently a research hotspot for new anti-infection and anti-tumor drugs. [0003] Alloferon-1 was first discovered in the blood of the red-heade...

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): C12N15/12C12N15/70C07K14/435A61K38/17A61P35/00
Inventor 孙琦孙爱华严杰
Owner 严杰
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