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Transcription factor of regulating and controlling vegetable fatty acid metabolism and its coding gene and application

A plant fatty acid and transcription factor technology, applied in a transcription factor regulating plant fatty acid metabolism and its coding gene and application field, can solve problems such as changing fatty acid content

Inactive Publication Date: 2007-04-18
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above research results show that fatty acid metabolism in plants is a complex and highly coordinated process, and the genetic manipulation of individual or single genes in the fatty acid metabolism pathway cannot effectively change the fatty acid content

Method used

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  • Transcription factor of regulating and controlling vegetable fatty acid metabolism and its coding gene and application
  • Transcription factor of regulating and controlling vegetable fatty acid metabolism and its coding gene and application
  • Transcription factor of regulating and controlling vegetable fatty acid metabolism and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the cloning of the transcription factor gene AtLEC1 that regulates plant fatty acid metabolism

[0056] Use TRIZAL reagent (purchased from Invitrogen Company) and refer to the kit instructions to extract the total RNA of fresh fruit pods of Arabidopsis thaliana (Arabidopsis thaliana) Columbia ecotype (Col-0), and then use Invitrogen Company's SuperScript TM II Reverse Transcriptase kit and refer to the kit instructions to synthesize the first strand cDNA by reverse transcription, and then use the synthesized cDNA as a template, in primer P1 (upstream primer): 5'-CCCTCGAGAAATGACCAGCTCAGTCGTAGT-3' and P2 (downstream primer) : Under the guidance of 5'-AAACTAGTTCACTTATACTGACCAT-3', PCR amplifies the transcription factor gene that regulates plant fatty acid metabolism in Arabidopsis thaliana. After the reaction, the PCR amplification product is detected by 1% agarose gel electrophoresis. The DNA recovery kit of Dingguo Company recovered and purified the target...

Embodiment 2

[0057] Embodiment 2, the acquisition of AtLEC1 transgenic Arabidopsis

[0058] 1. Construction of plant expression vectors containing AtLEC1

[0059] The plasmid pGEM-TVector-AtLEC1 containing AtLEC1 constructed in Example 1 was double digested with restriction endonucleases Xho I and Spe I, and the double digestion product was detected by 1% agarose gel electrophoresis, and the recovered length was about 717bp The AtLEC1 gene fragment was purified and the recovered fragment was purified with T 4 DNA ligase (Roche Company) and vector PER8 (Zuo et al., Plant Journal.24: 265-273, 2000) or pX6 (Zuo et al., Nature Biotechnology 19: 157-161, 2001) connection, then the connection product was transformed into Escherichia coli (E.coli) DH5α competent cell by heat shock method, screened positive clone, it was inoculated in the 5mL LB liquid medium containing 50mg / L hygromycin, at 37 Cultivate at 200rpm for 12-16 hours, extract the plasmid, and identify the recombinant plasmid with re...

Embodiment 3

[0064] Example 3, detection of fatty acid metabolism of AtLEC1 overexpression plants

[0065] 1. Sudan red staining indicates changes in fatty acid content in AtLEC1 overexpression plants

[0066] The AtLEC1 transgenic plants grown for 20 days after germination in the induction medium in Example 2 and the Columbia wild-type control plants were soaked in 1% Sudan Red (Fat Red 7B) (Sigma Company), room temperature overnight (12-24 hours), Wash with deionized water 3 times, each time for 60 seconds, the staining results are shown in Figure 4, the accumulation of fatty acids in the roots and cotyledons of AtLEC1 transgenic plants was significantly higher than that of the wild type.

[0067] 2. GC-MS method to detect the content changes of various fatty acid components in AtLEC1 transgenic plants after induction

[0068] Take respectively 0.1 g of the whole plants of the AtLEC1 transgenic plants and control plants obtained in Example 2 and grow for 20 days after germination in the...

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Abstract

This invention discloses a transcription factor and its coding gene and its application which regulating plant fatty acid metabolism. The purpose is to provide a transcription factor and its coding gene and its application in regulating plant fatty acid metabolism. The transcription factor is one of the following amino acid residue series. 1) SEQ ID NO1 in series table. 2) One to ten amino acid residue of SEQ ID NO:1 is displaced, absence or added to protein that can control plant fatty acid metabolism.

Description

technical field [0001] The invention relates to a transcription factor regulating plant fatty acid metabolism and its coding gene and application, in particular to a transcription factor regulating plant fatty acid metabolism and its coding gene derived from Arabidopsis thaliana and its application in regulating plant fatty acid metabolism. Background technique [0002] The main components of fats or vegetable oils in plants are fatty acids and their fatty acid derivatives, which are widely found in the seeds and fruits of plants. Vegetable oil is widely used, and the use of biotechnology to improve the composition and content of fatty acids in plants has important economic value. Among them, the operating gene selected by genetic engineering technology must be the key gene in the process of fatty acid biosynthesis in plants, but people still don't know much about the molecular regulation mechanism of plant fatty acid biosynthesis. Therefore, the use of genetic engineering t...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82
Inventor 左建儒牟金叶
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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