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

Escherichia coli engineering bacterium containing gene of Epl1 (eliciting plant response protein1) and construction method of escherichia coli engineering bacteria

A technology of Escherichia coli and construction methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as difficult to directly separate Epl1 protein, many proteins and metabolites, and heavy workload, and achieve an improvement Defense ability, wide application prospect, growth-promoting effect

Inactive Publication Date: 2014-06-18
NORTHEAST FORESTRY UNIVERSITY
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that there are many proteins and metabolites produced after the fermentation of Trichoderma dark green mycelium, it is difficult to directly separate the Epl1 protein, the workload is large, and the cost is high, and a strain containing a plant-stimulating response protein is provided. Escherichia coli engineering bacteria with Epl1 gene and its construction method

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
  • Escherichia coli engineering bacterium containing gene of Epl1 (eliciting plant response protein1) and construction method of escherichia coli engineering bacteria
  • Escherichia coli engineering bacterium containing gene of Epl1 (eliciting plant response protein1) and construction method of escherichia coli engineering bacteria
  • Escherichia coli engineering bacterium containing gene of Epl1 (eliciting plant response protein1) and construction method of escherichia coli engineering bacteria

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0031] Specific embodiment one: In this embodiment, an Escherichia coli engineering bacterium BL21-Epl1 containing the plant-stimulating response protein Epl1 gene is a Gram-negative short bacillus, a facultative anaerobic bacterium, and the colonies are round, smooth, colorless, and transparent .

[0032]The Escherichia coli engineered bacteria containing the stimulatory plant response protein Epl1 gene of this embodiment can inhibit the salicylic acid, jasmine Acid and auxin signaling pathway related gene expression can be significantly stimulated, and can cause significant changes in the activity of physiological enzymes in Populus japonicus. Therefore, the recombinant protein rEpl1 can not only induce the resistance of Populus japonicus, improve the defense ability of Populus japonicus, but also effectively promote the growth of Populus japonicus.

specific Embodiment approach 2

[0033] Specific embodiment two: the construction method of the Escherichia coli engineering bacterium that contains the stimulatory plant response protein Epl1 gene of this embodiment is carried out by the following steps:

[0034] 1. Extract the mycelium cDNA of Trichoderma dark green: at 28 ℃, under the condition of 200 rpm, culture the mycelium of Trichoderma dark green in 1 / 4PD medium for 48 hours, collect the mycelium, and then extract the total RNA, and reverse transcription of total RNA into cDNA;

[0035] Two, the construction of the prokaryotic expression vector pGEX-Epl1: the mycelia cDNA of Trichoderma dark green obtained in step 1 is used as a template, and Ep1e and Ep2e are primers for PCR amplification and purification to obtain a PCR product; then from Escherichia coli TOP10- Plasmid pGEX-4T-2 was extracted from pGEX-4T-2, and then the plasmid pGEX-4T-2 and the PCR product were first digested with BamHI, and the digested products were recovered, and then the dig...

specific Embodiment approach 3

[0040] Specific embodiment 3: The difference between this embodiment and specific embodiment 2 is that the PCR amplification reaction system described in step 2 is: 10×buffer 2 μL, dNTP 0.8 μL, template 2 μL, primer Ep1e1 μL, primer Ep2e1 μL, deionized water 12.8 μL, EXTaq 0.4 μL, total volume 20 μL. Reaction program: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30s, annealing at 58°C for 40s, extension at 72°C for 80s, 35 cycles; fill-in at 72°C for 20 minutes, and storage at 4°C, where the template is cDNA, primer Ep1e and primer Ep2e The concentration is 20 μM. Others are the same as in the second embodiment.

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 an escherichia coli engineering bacterium containing the gene of Epl1 (eliciting plant response protein1) and a construction method of the escherichia coli engineering bacterium.. The escherichia coli engineering bacterium BL21-Epl1 containing the gene of Epl1 is gram negative brevibacterium and facultative anaerobe, and the bacterial colony of the escherichia coli engineering bacterium BL21-Epl1 is circular, smooth, colorless and transparent. The construction method comprises the following steps: I, extracting a mycelium cDNA (Complementary Deoxyribonucleic Acid) of trichoderma atroviride; II, constructing a prokaryotic expression vector Pgex-EPl1; and III, converting escherichia coli to obtain the escherichia coli engineering bacterium. Under the condition that the fermenting property is not affected, after being induced by a recombinant protein rEPl1, the escherichia coli engineering bacterium provided by the invention remarkably excites the expression of genes related to signal transfer paths of salicylic acid, jasmonic acid and auxin signals of Shanxinyang. The production process of the escherichia coli engineering bacterium is simple and low in cost. The escherichia coli engineering bacterium containing the gene of Epl1 (eliciting plant response protein1) and the construction method thereof are applied to the agricultural field.

Description

technical field [0001] The invention relates to an Escherichia coli engineering bacterium containing a plant-stimulating response protein Epl1 gene and a construction method thereof. Background technique [0002] Many studies have found that the biological control of plant fungal diseases Trichoderma atroviride can stimulate the plant's own defense system to enable plants to resist the invasion of pathogens, that is, induce plant disease resistance, thereby promoting plant growth. The mechanism of Trichoderma-induced plant disease resistance is the overall improvement of plant immunity, while the mechanism of competition, parasitism, and resistance is a single process of action between plant disease biological control bacteria and pathogenic microorganisms. Systemic acquired resistance (SAR) and induced systemic resistance (Induced systemic resistance, ISR) are two forms of induced resistance. SAR refers to the broad-spectrum, long-lasting and systemic resistance of plants ...

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): C12N1/21C12N15/70C12R1/19
Inventor 刘志华遇文婧王志英刁桂萍张荣沭范海娟黄颖宋小双
Owner NORTHEAST FORESTRY UNIVERSITY
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