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

Method for improving artemisinin content in Artemisia annua L. by DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene transfer

A kind of artemisinin and transgenic technology, which is applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of difficult artificial synthesis, complex structure of artemisinin, and infeasibility, and achieve stable The effect of new drug sources

Inactive Publication Date: 2012-07-25
SHANGHAI JIAO TONG UNIV
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the main source of artemisinin is extracted from the aerial parts of Artemisia annua plants. However, the content of artemisinin in Artemisia annua is very low. The average content of artemisinin in different planting environments and planting varieties is 0.01-1% of the dry weight of leaves of Artemisia annua. , making the large-scale commercial production of this drug limited
Due to the complex structure of artemisinin, the artificial synthesis is difficult, the yield is low, and the cost is high, so it is not feasible
Some people try to produce artemisinin by tissue culture and cell engineering. However, the content of artemisinin in callus is less than 0.1% of dry weight, and the highest in shoot is only 0.16% of dry weight. Artemisinin was not detected in roots
Therefore, the feasibility of using tissue culture and cell engineering to produce artemisinin is not high.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0027] Step 1, cloning of Artemisia annua DXR gene

[0028] (1) Extraction of total RNA from Artemisia annua genome

[0029] Take 100-200 mg young leaves of Artemisia annua, quick-freeze them with liquid nitrogen, grind them quickly with a mortar, add them to a 1.5 mL Eppendorf tube filled with 1 mL TRlzol (TRlzol Reagents, GIBCOBRL, USA), shake them fully, and place them at room temperature Leave for 5 minutes, add 200 μL of chloroform, shake vigorously for 15 sec, place at room temperature for 2-3 minutes, and centrifuge at 12,000 rpm for 15 minutes at 4°C; suck the supernatant (about 600 μL) into a clean 1.5 mL Eppendorf tube, add an equal volume of isopropyl Alcohol, mix by inverting, place at room temperature for 10min, then centrifuge at 12,000rmp for 10min at 4°C; discard the supernatant, add 1mL of 75% ethanol to wash, shake, and centrifuge at 7,500rmp for 5min at 4°C; dry at room temperature for 10-15min Dissolve in an appropriate amount (30-40μL) of RNAase-free wate...

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 provides a method for improving an artemisinin content in Artemisia annua L. by DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene transfer. The method comprises the following steps of: cloning DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene in the Artemisia annua L., so as to establish a plant expression carrier containing the DXR gene; transferring the DXR gene into Artemisia annua L. through agrobacterium tumefaciens-mediated transformation to grow a plant; detecting the integration condition of the exogenous target gene DXR by a PCR (Polymerase Chain Reaction); detecting the artemisinin content in the transgenic Artemisia annua L. through high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD); and screening the transgenic Artemisia annua L. plant with improved artemisinin content. The artemisinin content of the transgenic Artemisia annua L. obtained by the invention is obviously improved and high up to 2.4 times that of the non-transferred plant. Therefore, the invention provides the method for improving the artemisinin content in the Artemisia annua L. and lays a foundation for producing artemisinin by using the transgenic Artemisia annua L. in a large scale.

Description

technical field [0001] The invention relates to a method for increasing the content of artemisinin in the field of biotechnology, in particular to a method for increasing the content of artemisinin in Artemisia annua by transducing DXR gene. Background technique [0002] Artemisia annua L. is an annual herb belonging to the family Asteraceae. Artemisinin (artemisinin) is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is currently the most effective drug for treating malaria in the world, especially for cerebral malaria and anti-malaria Chloroquine malaria has the characteristics of quick action and low toxicity. Currently, the most effective treatment for malaria recommended by the World Health Organization is artemisinin combination therapy (ACTs). In addition, with the deepening of pharmacological research on artemisinin, scientists have discovered that artemisinin and its derivatives also have anti-inflammatory,...

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): C12N15/84A01H5/00
Inventor 唐克轩王玥月陈韵斐吴韶龑王涛沈乾
Owner SHANGHAI JIAO TONG UNIV
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