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

Method for increasing proanthocyanidin content in escherichia coli by cotransformation of brassica juncea gene ANS and brassica carinata BAN

A technology of Brassica napus, Escherichia coli, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc.

Inactive Publication Date: 2012-12-19
HUNAN UNIV OF SCI & TECH
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching the prior art literature, it was found that YanY J et al. (2008) reported on the "Biotechnol. Bioeng. 2008; 100: 126-140" by expressing Arabidopsis and Gerbera in Escherichia coli (Gerbera), Petunia (Petunia), Antirrhinum (Anthocyanin) synthesis pathway genes can increase the synthesis of E. coli anthocyanin, anthocyanin and proanthocyanin are flavonoids , but there is no report on using the proanthocyanidin synthesis gene of Brassica species to increase the synthesis of E. coli proanthocyanidin

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 Construction

[0016] The embodiments of the present invention are described in detail below: the present embodiment implements under the premise of the technical solution of the present invention, provides detailed implementation and specific operation process, but protection scope of the present invention is not limited to the following implementation .

[0017] The experimental method that does not indicate concrete condition in the embodiment, generally according to routine condition, for example " molecular cloning: laboratory handbook " (New York: Cold Spring Harbor Laboratory Press, 1989) such as Sambrook, " modern molecular biology such as Zheng Weijuan Experiment" (Beijing: Higher Education Press, 2010), or according to the conditions suggested by the manufacturer.

[0018] (1) Cloning of rapeseed ANS and BAN genes

[0019] (1) Extraction of total RNA from mustard rape seeds and Ethiopian mustard seeds

[0020] A small amount of seeds 20-25 days after pollination of mustard type r...

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 increasing proanthocyanidin content in escherichia coli by cotransformation of brassica juncea gene ANS and brassica carinata BAN. According to the invention, an anthocyanidin synthetase ANS gene is cloned from cDNA of brassica juncea seeds, and an anthocyanidin reductase BAN gene is cloned from cDNA of brassica carinata seeds; a prokaryotic expression vector pET-ANS-BAN containing the ANS and BAN genes is constructed; and the ANS and the BAN genes are introduced into a BL21 escherichia coli strain simultaneously. The transformed strain is cultured in an LB liquid medium at 28-38 DEG C at 150-250 rpm for 6-12 hours, transferred into an LB liquid medium containing grape skin extracts (0.2-0.8 g / L), and cultured under shaking at 28-38 DEG C at 150-250 rpm for 36-54 hours; the strain is collected by centrifugation; and the proanthocyanidin content in escherichia coli is determined by a DMACA-HCl method. The transgenic escherichia coli obtained in the invention has significantly increased proanthocyanidin content, and the maximum content can be 24.2 times higher than that of a non-transformed control strain.

Description

technical field [0001] The present invention relates to a method in the field of genetic engineering and microbial fermentation technology, in particular to a method for co-transformation of double key enzyme genes and adding crude extracts of key enzyme gene catalyzed reaction precursors in the culture medium to improve Escherichia coli Method for synthesizing proanthocyanidins. Background technique [0002] Proanthocyanidins (Proathocyanidins), also known as proanthocyanidins, are a type of flavonoids. Proanthocyanidin is one of the strongest antioxidants for scavenging free radicals, which can protect brain and nerve tissue, improve blood circulation, flexible joints and youthful skin. Long-term consumption of foods high in proanthocyanidins can alleviate cardiovascular and other diseases. Scientists have also discovered that proanthocyanidins and their derivatives also have anti-inflammatory, anti-adversity, anti-tumor, and immune-regulating functions. It can be seen t...

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): C12N15/70C12N1/21C12R1/19
Inventor 刘丽莉严明理向建华孙婵尹小明
Owner HUNAN UNIV OF SCI & TECH
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