Unlock instant, AI-driven research and patent intelligence for your innovation.

Chamomile farnesyl diphosphatesynthase gene clone and prokaryotic expression

A technology of farnesyl pyrophosphate and chamomile, applied in genetic engineering, plant gene improvement, transferase, etc., can solve problems that have not yet been seen

Inactive Publication Date: 2012-10-03
ANHUI AGRICULTURAL UNIVERSITY
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are no research reports on key enzyme genes such as FPS gene in the metabolic synthesis pathway of chamomile terpenoids at home and abroad.

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
  • Chamomile farnesyl diphosphatesynthase gene clone and prokaryotic expression
  • Chamomile farnesyl diphosphatesynthase gene clone and prokaryotic expression
  • Chamomile farnesyl diphosphatesynthase gene clone and prokaryotic expression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0037] 1 Materials and methods

[0038] 1.1 Materials

[0039]Chamomile (Matricaria chamomilla L.) was provided by Xuancheng Yueping Ecological Co., Ltd. Escherichia coli DH5α, BL21(DE3), and prokaryotic expression vector pET-30a (+) were provided by the College of Life Sciences, Anhui Agricultural University. The cloning vector PMD19-T was provided by TAKARA Company. Restriction enzyme, DNA polymerase, T4 ligase, IPTG (isopropyl-β-D-thiogalactopyranoside), X-gal (5-bromo-4-chloro-3-indole- β-D galactoside), DNA gel recovery kit, plasmid small extraction kit, RNAiso plus (total RNA extraction reagent), etc. were purchased from TakaRa Company. Gene sequencing was completed by Shanghai Handsome Biotech Co., Ltd.

[0040] 1.2 Method

[0041] 1.2.1 Chamomile total RNA extraction.

[0042] (1) Grind 0.2g of chamomile petals with liquid nitrogen to powder

[0043] (2) Add 2ml of total RNA extraction reagent, cover the sample, and let it stand at room temperature for 15 minute...

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 chamomile farnesyl diphosphatesynthase and a construction and expression method of a recombinant expression vector thereof, the amino acid sequence of the chamomile farnesyl diphosphatesynthase is shown as SEQ NO2, and the nucleotide sequence of the chamomile farnesyl diphosphatesynthase is shown as SEQ NO1. The ORF (open reading frame) of the chamomile FPS (frames per second) gene is 1032bp, and the chamomile FPS gene has higher homology compared with other plant sesquiterpene synthase genes through sequence comparison; and Westerning verification can be carried out by constructing a prokaryotic expression vector pET-30a(+) / FPS and inducting for expressing soluble protein to obtain a target protein with molecular weight of 38KDa, so that a foundation is established to further studies on FPS protein purification and protein structure and characteristics, and a reference is provided for improving the yield of chamomile chamazulene by a transgene method. Furthermore, the condition for induction expressing of protein is optimized in the method, the optimal induction time is six hours, and the optimal induction temperature is 37 DEG C.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a chamomile farnesyl pyrophosphate synthase gene cloning, prokaryotic expression and application. Background technique [0002] Chamomile (Matricaria chamomilla L.) is a herbaceous plant of the genus Matricaria in the Asteraceae family. The whole plant is fragrant and contains volatile aromatic oils. Its volatile oil has anti-inflammatory, anti-fungal, antispasmodic, sedative, pain-relieving and improving symptoms of lung cancer patients. Its essential oil is widely used in food, beverage, tobacco, daily chemical, medicine and other fields due to its unique aroma and antioxidant effect. It is a new variety of agricultural industrial structure adjustment with great market potential, which integrates landscaping, beautification, fragrance and color. [0003] The active ingredients in chamomile are mainly concentrated in volatile oil, and the volatile oil content in flowers is general...

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
IPC IPC(8): C12N9/10C12N15/54C12N15/70C12N15/66
Inventor 袁艺邰玉玲杨秀梅
Owner ANHUI AGRICULTURAL UNIVERSITY