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

New 0 - ultra family conantokins, coded polynucleotide and application

A technology of conotoxin peptides and polynucleotides, applied in the biological field, can solve problems such as insufficient in-depth research

Pending Publication Date: 2006-07-05
HAINAN UNIVERSITY
View PDF0 Cites 25 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although several conotoxin peptides of the O superfamily have been discovered, few of them have been studied in depth

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
  • New 0 - ultra family conantokins, coded polynucleotide and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Cloning of O-superfamily conotoxin gene

[0077] 1 Extraction of total RNA from conus snails

[0078] Conus textile Linnaeus, Conus betulinus Linnaeus, Conus lividus Hwass, Conus littertus Linnaeus and Warrior conus collected from the coasts of Hainan Island and Xisha Islands. (Conus miles Linnaeus), Conus caracteristicus Fischer, Conus vexillum Gmelin, Conus capitaneus Linnaeus, Conus marmoreus Linnaeus, Conus magus L., Conus virgo L. and Conus emaciatus Reeve, a total of 12 live conus species, were used as materials. Total RNA was extracted according to the operating manual with a small column centrifugal tissue / cell total RNA extraction kit (Shanghai Huasun Bioengineering Co., Ltd.), or with a Total RNA IsolationSystem kit (Promega). Let's take the RNA extraction kit produced by Shanghai Huasun as an example.

[0079]First, freeze the cone venom gland or poisonous tube with liquid nitrogen and grind it into powder, and transfer it to a 1.5mL centrifuge t...

Embodiment 2

[0097] Example 2 Synthesis of O-superfamily conotoxins MiEr93M (SEQ ID NO: 10), MgJr93M (SEQ ID NO: 14) and CaFr179M (SEQ ID NO: 33)

[0098] According to the amino acid sequences of conotoxin peptides MiEr93M, MgJr93M and CaFr179M, these three peptides were artificially synthesized by Fmoc method. The specific method is as follows.

[0099] The above three conotoxin peptides were synthesized on an ABI Prism 433a peptide synthesizer by using the Fmoc method in the solid-phase synthesis method. The side chain protecting groups of Fmoc amino acids are: Pmc (Arg), Trt (Cys), But (Thr, Ser, Tyr), OBut (Asp), Boc (Lys). Using Fmoc HOBT DCC method, Rink resin and Fmoc amino acids, The synthesis steps were carried out referring to the synthesis manual of the instrument. In order to complete the reaction, the piperidine deprotection and coupling time were appropriately extended, and double coupling was used for difficult amino acids. The crude linear peptide was recovered and purif...

Embodiment 3

[0100] Example 3 Oxidative folding of conotoxins MiEr93M, MgJr93M and CaFr179M linear peptides

[0101] Purified reduced linear peptides MiEr93M, MgJr93M and CaFr179M were oxidatively folded at room temperature for 24h, respectively. The oxidative folding buffer consisted of: 0.1 M ammonium acetate (pH 7.8), 20 μM fully reduced linear peptide, reduced (GSH) and oxidized (GSSG) glutathione. The concentration ratio of peptide:GSH:GSSG in the buffer was 1:50:5. At the end of the oxidation reaction, the pH was adjusted to 23 with TFA (trifluoroacetic acid) to terminate the oxidation reaction. Peptides were purified using a liquid chromatography system (Bio-Rad). The reaction mixture was loaded on a preparative chromatography column (4.6×250 mm, Sinochrom ODS-BP) at a flow rate of 1 ml / min, and eluted with 0.1% TFA until all the reaction solution was eluted completely. Solvent A was 0.1% TFA in acetonitrile and solvent B was 0.1% TFA in water. Gradient elution (10% → 100% A, 90...

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

This invention describes a new O-Conotoxin (O-CTX), its encoding polynucleotide, the construction units and transformation cells having this encoding polynucleotide, and the reconstruction process for producing the said O-Conotoxin. This invention also describes the artificial synthesis and applications of the said O-Conotoxin.

Description

technical field [0001] The invention relates to a new O-superfamily conotoxin peptide, its encoding polynucleotide and its use, and belongs to the field of biotechnology. Background technique [0002] Conotoxin (CTX) is a small peptide toxin secreted by Conus, a carnivorous mollusc living in tropical oceans, used to anesthetize prey. They are composed of 6-41 amino acid residues and are rich in cysteine ​​(Cys) animal neuropeptide toxins. It is extremely toxic and can cause convulsions, tremors, and even paralysis in animals. The conotoxin has a novel chemical structure, strong biological activity, and high selectivity of the target. It can be directly used as a natural medicine or as a lead compound for drug design. In view of its small molecule, stable structure, large range of action on receptors, strong specificity, and chemical synthesis; it is also the product of gene expression, and it can also be recombinantly expressed in vitro to obtain a large amount of toxin. ...

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): C07K14/435C12N15/12C12N15/11C12N15/63A61K38/17A01N37/00A61P29/00
Inventor 罗素兰长孙东亭张本林秋金
Owner HAINAN 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