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

Recombinant plasmid containing PON gene and its use

A technology of recombinant plasmids and genes, applied in the field of biomedicine, which can solve the problems of host immune and inflammatory response, prone to mutation or random integration, etc.

Inactive Publication Date: 2013-04-17
INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Viral vectors can be highly expressed in vivo, but they are prone to mutation or random integration, triggering severe immune and inflammatory responses and other unpredictable serious consequences in the host

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
  • Recombinant plasmid containing PON gene and its use
  • Recombinant plasmid containing PON gene and its use
  • Recombinant plasmid containing PON gene and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Construction of recombinant plasmid vector

[0035] The prokaryotic expression vector pBluescrip / PON (gifted by Dr. Clement E. Furlong) containing the human PON gene (see SEQ ID NO: 1 sequence) was used as a template to design primers: primer 1, 5'GCA GGTACC ATGGCGAAGCTGATTGCG 3′; Primer 2, GCA GAATTC TTAGAGCTCACAGTAAAG 3'. KpnI and EcoRI restriction sites were inserted into primer 1 and primer 2, respectively. Perform PCR amplification under 40 cycles of 94C for 40 seconds, 58C for 40 seconds, and 72C for 2.5 minutes. The amplified product was digested with KpnI and EcoRI (under the action of T4 ligase) and inserted into pDNA, and sequenced after PCR and enzyme digestion identification were correct to obtain the pDNA / PON plasmid ( figure 1 ). The plasmid was amplified in Escherichia coli TOP10 strain, and the plasmid was extracted.

[0036] In addition, according to the PON gene sequence disclosed herein or other sequences, those skilled in the art can isolate o...

Embodiment 2

[0038] Prevention of soman poisoning by pDNA / PON

[0039] The organophosphorus compound soman is a highly toxic military agent and can also be used in terrorist activities. Its toxicity is mainly manifested as the irreversible inhibition of cholinesterase, which phosphorylates the serine residue in the enzyme active site of cholinesterase, leading to the inactivation of the enzyme and the accumulation of acetylcholine in the nerve endings. PON can catalyze the hydrolysis of the phosphoester bond in the soman molecule and protect against a certain degree of soman poisoning (Table 1). In this experiment, the plasmid pcDNA / PON was directly injected intravenously into the mice, and 200 μg / kg (1.0 LD) of sc Soman was given to the mice 24 hours later. The normal control and mice injected with empty plasmids showed muscle tremors, vertical tail, tremor, and salivation. , convulsions, and died within about 9 minutes, while 60% of mice pre-injected with 600 μg pcDNA / PON survived (obse...

Embodiment 3

[0041] pDNA / PON prevents atherosclerosis

[0042] The surface of normal vascular intima is smooth without lipid infiltration. After feeding with high-fat diet for 25 days, the aortic intima had serious pathological changes, the surface was rough, and lipid plaques were formed higher than the surface of the intima, especially in the aortic root, where obvious lipid plaques appeared. It shows that pathological changes of atherosclerosis have been formed. If given (tail vein injection) model rats pcDNA / PON for 125 days at the same time, the plaques on the wall of the aorta decreased or disappeared, and the pathological changes were greatly improved ( figure 2 ). It shows that the recombinant plasmid genome can be effectively expressed in mice, and the PON in the expression product has played an antioxidant role in the serum, increasing the level of SOD, reducing the production of lipid peroxidation products, and reducing serum TG, LDL-C And TC, remove fat accumulation in 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 is about the recombination plasmid containing the PON gene and its application. The recombination plasmid, the cell and the compound can prevent and cure the toxic of the organophosphor, hyperlipemia, atherosclerosis, glycuresis.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of the recombinant plasmid carrying serum paraoxonase gene in the prevention and treatment of organophosphorus compound poisoning, hyperlipidemia, atherosclerosis, diabetes and its complications. Background technique [0002] Serum paraoxonase (PON) is a calcium ion-dependent aromatic esterase secreted by the liver. There are three members of the PON gene family, including PON1, PON2, and PON3, which are located on chromosome 7q21.3-22.1. There is a hydrophobic region at the N-terminus of PON protein, and PON is closely combined with apolipoprotein in high-density lipoprotein through its hydrophobic sequence, and exerts various biological effects. The substrates of PON are aromatic esters, organic phosphates, and phospholipids of oxidized low-density lipoprotein. In patients with hyperlipidemia, atherosclerosis, and diabetes, the serum PON level and activity decrease,...

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 Patents(China)
IPC IPC(8): C12N15/63C12N15/52C12N15/79A61K48/00A61K38/43A61P3/06A61P3/10A61P9/10
Inventor 付爱玲孙曼霁
Owner INST OF PHARMACOLOGY & TOXICOLOGY ACAD OF MILITARY MEDICAL SCI P L A
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