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

Brassica carinata A. braun fatty acid elongase and coding gene thereof

A technology of fatty acid elongation enzyme and coding gene, which is applied in the fields of molecular biology and genetic engineering, and can solve problems such as the complex content of erucic acid

Inactive Publication Date: 2013-03-20
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The FAE1 (Fatty Acid Elongase 1) gene is a key gene that regulates erucic acid synthesis. Although James et al. first cloned the FAE1 gene from Arabidopsis thaliana, complete FAE1 genes have been isolated from various cruciferous plants. However, at present, the cloning of FAE1 gene is limited to a few plants of Arabidopsis and Brassica, and Brassicaceae is the main germplasm resource of erucic acid distribution. There are many types of plants and complex erucic acid content. Suitable materials for gene cloning

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
  • Brassica carinata A. braun fatty acid elongase and coding gene thereof
  • Brassica carinata A. braun fatty acid elongase and coding gene thereof
  • Brassica carinata A. braun fatty acid elongase and coding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1, the cloning of gene BcFAE1

[0017] To clone BcFAE1, the specific steps are as follows:

[0018] 1. Acquisition of BcFAE1 gene

[0019] Ethiopian mustard is cultivated under normal conditions. After the true leaves grow, the leaves are collected, DNA is extracted from the leaves, polymerase chain reaction is carried out, and the BcFAE1 gene is finally obtained.

[0020] 1. Extraction of Ethiopian thaliana genomic DNA

[0021] Take about 0.2g of fresh young living plant leaves, fully grind them under liquid nitrogen freezing conditions, transfer them to a 1.5mL centrifuge tube, add 500μL CTAB extract, and bathe in water at 65°C for 45min, during which time they are inverted and mixed 3 times; add 500μL volume Chloroform-isoamyl alcohol with a ratio of 24:1, mixed evenly, 12000r·min -1 Centrifuge for 15 min; take the supernatant, add 2 times the volume of absolute ethanol, and put it in the refrigerator at -20°C overnight; 4000r min -1 Centrifuge for 15m...

Embodiment 2

[0031] Example 2, Functional Verification of BcFAE1 Gene

[0032] 1. High expression of BcFAE1 gene in yeast

[0033] In order to demonstrate the coding function of the BcFAE1 gene, the present invention clones the BcFAE1 gene between the BamHI and KpnI sites of pYES2 / NT C from Invitrogen, and transforms the yeast strain InvSc1 to obtain high-level expression.

[0034] The present invention utilizes the multiple cloning sites of the pYES2 / NT C high-efficiency expression vector of Invitrogen Company to realize the high-efficiency expression of BcFAE1. The enzyme cleavage site used in this experiment makes the 5' end of the product expressed by the pYES2 / NT C expression vector add 6 consecutive His fusion proteins, and these 6 consecutive His constitute the specific binding site for i-NTA gel. Therefore, the expression product can be purified by affinity chromatography.

[0035] The pMD-BcFAE1 recombinant vector carrying the BcFAE1 gene and the pYES2 / NT C expression vector wer...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention provides a Brassica carinata A. braun fatty acid elongase and a coding gene thereof (BcFAE1). The gene is introduced in to yeast; and the erucic acid content in the recombinant yeast reaches 0.27+ / -0.24%. The protein and the coding gene thereof have important theoretical and practical significance for the research of a genetic control mechanism of plant erucic acids, the increasement of the erucic acid content and the improvement of related traits, can play an important role in the genetic engineering improvement of the plant erucic acid genes, and have wide application prospects.

Description

technical field [0001] The present invention relates to the field of molecular biology and genetic engineering, more specifically, relates to Ethiopian thalli (Brassica carinata A. Braun) fatty acid elongase (FAE1) and its coding gene. Background technique [0002] Erucic acid (C22:1) is a very long chain unsaturated fatty acid, mainly found in the seeds of Brassicaceae and Tropaeolaceae. As a metabolite of plants, erucic acid also affects the growth and development of plants to a certain extent, and participates in the synthesis of biofilm cutin or wax in the interaction between plants and the environment. In terms of nutritional value, erucic acid is an anti-nutritional component of edible oil, which is not easy to be digested and absorbed by the human body, and it is easy to cause diseases such as coronary heart disease and fatty liver. However, in industry, erucic acid and its derivatives, such as erucic acid amide, have good plasticity and hydrophobicity, so they have ...

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): C12N9/00C12N15/52C12N15/63C12N5/10C12N1/19C12N15/09A01H5/00
Inventor 李密密杭悦宇孙小芹庞慧李莹郭建林严琴琴
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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