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

Production and use of high expression, high purity, high activity gene recombinant human lysozyme

A technology of human lysozyme and genetic recombination, which is applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., to achieve high purity and high activity

Inactive Publication Date: 2006-12-13
长春奇龙生物技术研究所 +2
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methicillin-resistant Staphylococcus aureus (MRSA) has no cure other than vancomycin

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
  • Production and use of high expression, high purity, high activity gene recombinant human lysozyme
  • Production and use of high expression, high purity, high activity gene recombinant human lysozyme
  • Production and use of high expression, high purity, high activity gene recombinant human lysozyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Embodiment 1: the production method of the human lysozyme expressed by genetic engineering;

[0011] Materials: The expression system of genetically engineered human lysozyme is characterized by using methanolotrophic yeast as the host strain "Pichia genus strain". In the phenotype of methanol utilization, both fast methanol utilization (mut+) and slow methanol utilization (muts or mut-) were used. The smd1165(his4.prb1), smd1163(his.pep4.prb1), Gs115 and km71 strains of Pichia pastoris were mainly used. The plasmid of the genetically engineered human lysozyme of the present invention is mainly integrated into the chromosome of the above bacterial strain by using the plasmid of the pPIC9k series.

[0012] Steps: 1. Shake flask seed preparation: based on the preparation of 200 milliliters of culture medium, use H 3 PO 4 (phosphoric acid) 6 ml, MgSO 4 (magnesium sulfate) 3 grams, K 2 SO 4 (potassium sulfate) 4 grams, KOH (potassium hydroxide) 1 gram, CaSO 4 2H 2 O...

Embodiment 2

[0014] Embodiment 2: the identification of human lysozyme bacteriolytic activity (test tube method)

[0015] Material

[0016] 1. Buffer pH7.210mMTris-Cl buffer

[0017] 2. Strains Staphylococcus aureus, Escherichia coli

[0018] 3. 10% SDS

[0019] 4. LB medium

[0020] method:

[0021]

Embodiment 3

[0022] Example 3: The bactericidal effect of human lysozyme on drug-resistant bacteria, as shown in Figures 6 and 7, with Bacillus abalone and Staphylococcus aureus as examples.

[0023] Drug-resistant strains: 30 strains of different drug-resistant bacteria came from the Laboratory Department of Xi'an Xijing Hospital Method:

[0024] Prepare 1% agarose with pH7.2 10mM Tris-Cl buffer

[0025] After autoclaving, cool to 40 °C and add bacteria to 10 6 dump flat

[0026] ↓

[0027] Punch holes in the plate with a hole punch

[0028] ↓

[0029] Add different concentrations of lysozyme solution (0-100mg / ml) to the wells

[0030] ↓

[0031] Incubate in a 37°C incubator for 4-6 hours

[0032] ↓

[0033] After LB nutrient agar is autoclaved, cool to 40°C and cover on the above agarose plate

[0034] ...

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

A high-expression, high-purity and high-activity gene-recombinant human lysozyme, its preparing process and the medical composition of antibiotic and the said human lysozyme for treating bacterial and fungus infection, drug-resistant bacteria infection, and the sequela caused by gram-negative bacteria infection are disclosed.

Description

Technical field: [0001] The invention relates to a method and application of gene recombinant human lysozyme, in particular to the production and application of a gene recombinant human lysozyme with high expression, high purity and high activity, belonging to the technical field of bioengineering. Background technique: [0002] Lysozyme was discovered by Fleming in 1922 (Proc. R. Soc. Lond. B. Biol. Sci. 1922). It is an effective antibacterial agent, its full name is: 1,4-β-N-lysozyme or: mucopeptide N-acetylmuramoyl hydrolase. It can cut off the β-1,4 glycosidic linkage between N-acetylglucosamine and N-acetylmuramic acid in the peptidoglycan of the bacterial cell wall, and destroy the peptidoglycan scaffold. Swells and cracks, causing bacteria to lyse. Human and animal cells have no cell wall structure and no peptidoglycan, so lysozyme has no toxic effect on human cells. Lysozyme widely exists in animals, plants and microorganisms in nature. At present, people mainly ...

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): C12N15/81C12N1/19C12N9/36C12N15/56A61K38/47A61P31/00A61P31/04
Inventor 安米张华李元安献禄
Owner 长春奇龙生物技术研究所
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