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

A kind of method for producing mycelomycin with Artemisia annua

A technology of mycelia and Artemisia annua, applied in the fields of biotechnology and genetic engineering, can solve problems such as plasmid instability, decreased expression of exogenous mycelia gene, and decreased copy number

Active Publication Date: 2018-12-25
CHANGSHA ZHONGKEJINGBO BIOTECH
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, once the laboratory is expanded to industrial scale production, due to the disappearance of the selection pressure to maintain a high copy number of the plasmid in the culture medium, the plasmid becomes unstable and the copy number decreases, so the expression level of the exogenous plectasin gene also increases. decline

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
  • A kind of method for producing mycelomycin with Artemisia annua
  • A kind of method for producing mycelomycin with Artemisia annua
  • A kind of method for producing mycelomycin with Artemisia annua

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0119] Specific embodiments of the invention The sequences used in the examples are the sequences mentioned above

[0120] The method for producing Plectasin with Artemisia annua of the present invention, it comprises the following steps:

[0121] a. Transformation carrier T202;

[0122] b. Constructing the recombinant plasmid T202-Plec expressing plectasin gene in Artemisia annua cells;

[0123] c. Transform Artemisia annua with recombinant plasmid T202-Plec;

[0124] d. Screen positive transgenic Artemisia annua seedlings, and carry out real-time RT-PCR identification.

[0125] The specific operation process is:

[0126] 1. Transformation carrier T202

[0127] Clone the pRB fragment:

[0128] Using pCambia1300 as a template, design synthetic primers to amplify pRB ( figure 1 ). PCR reaction conditions: pre-denaturation at 94°C for 5 minutes, 30 cycles at 94°C for 30s-59°C for 30s-72°C for 70s, and 72°C for 7 minutes. After 2% agarose gel electrophoresis, the target ...

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 method for producing plectasin through sweet wormwood herbs. The method includes the following steps of firstly, refitting and obtaining a carrier T202; secondly, optimizing and amplifying a plectasin gene; thirdly, establishing recombinant plasmid T202-Plec for expressing the plectasin gene in sweet wormwood herb cells; fourthly, converting agrobacterium EHA105 cells through the recombinant plasmid T202-Plec; fifthly, screening out positive strains, and conducting enzymatic detection; sixthly, transforming the sweet wormwood herbs to obtain sweet wormwood herb transformation plants with positive antimicrobial peptide plectasin; seventhly, quantitatively authenticating the expression amount of plectasin through PCR(qRT-PCR) in real time. An sweet wormwood herb expression system adopted for the method has the advantages that according to the codon preference of the sweet wormwood herbs, the nucleotide sequence of the plectasin gene is optimized, and the efficient and stable mRNA transcription, rapid and accurate translation and high external protein accumulation are ensured; plant expression plectasin is low in cost, safe and high in economic benefit; the expressed protein product can be directly used for being added without being purified.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and genetic engineering, in particular to a method for producing plectasin by using Artemisia annua. Background technique [0002] China is a large animal husbandry country, and the animal feed produced every year suffers huge losses due to corruption and deterioration during storage. Therefore, a large number of manufacturers add chemical preservatives to feed, but the chemical preservatives left in the animal body not only affect the quality of livestock and poultry products, but also threaten people's health, such as causing gastroenteritis and other diseases, affecting liver and kidney function etc. On the other hand, in order to prevent animal husbandry diseases, antibiotics have been added to animal feed in large quantities for a long time, and the problems of animal digestive tract disorders and drug resistance caused by this are also becoming more and more serious. [0003] Antiba...

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): C12N15/82A01H5/00A01H6/14
Inventor 夏新界崔延春余艳尹晟
Owner CHANGSHA ZHONGKEJINGBO BIOTECH
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