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A kind of anti-oxidation and high-salt stress-resistant artificially synthesized srna and its application

An artificial synthesis and anti-oxidation technology, applied in the field of genetic engineering, can solve the problems of undetermined coding genes and functions, no anti-oxidation and high-salt stress resistance, no host adversity stress, etc., to achieve the improvement of oxidative stress Effects of resistance and salt stress resistance, high survival

Active Publication Date: 2017-12-15
北京绿氮生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] sRNA was first discovered in E. coli forty years ago, but its coding gene and function have not been determined
[0004] However, there has not been any research report about the non-coding sRNA gene from nitrogen-fixing Pseudomonas stutzeri endowing host cells with the functions of anti-oxidation and high-salt stress resistance, let alone that it can improve host adversity stress by binding to the Hfq transcriptional regulator. sex reports

Method used

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  • A kind of anti-oxidation and high-salt stress-resistant artificially synthesized srna and its application
  • A kind of anti-oxidation and high-salt stress-resistant artificially synthesized srna and its application
  • A kind of anti-oxidation and high-salt stress-resistant artificially synthesized srna and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Determination of specific binding sequence in artificially synthesized sRNA and construction of expression plasmid pSAncR03

[0034] 1. Determination of the specific binding sequence in artificially synthesized sRNA

[0035] We found that non-coding sRNA03 from nitrogen-fixing Pseudomonas stutzeri A1501 was involved in regulating some stress-resistant physiological processes of host cells. Experiments have proved that the lack of sRNA03 will lead to the reduction of the antioxidant and salt tolerance of cells, and the transfer of this sRNA into E. coli will also significantly enhance its survival in peroxidative and high-salt environments, which regulates the physiological process of cell stress resistance has been confirmed. In order to construct sRNA with stronger regulatory ability, the specific binding sequence needs to be clarified first. For this reason, we analyzed sRNA03 through bioinformatics software and selected a 20bp fragment of its binding regio...

Embodiment 2

[0050] Example 2 Construction of mutant strains and verification of their functions

[0051] A method for improving oxidative stress resistance and / or salt stress resistance of Escherichia coli TOP10 strain by artificially synthesizing sRNA is used to construct mutant strains with oxidative stress resistance and / or salt stress resistance.

[0052] In this embodiment, the artificially synthesized sRNA gene is a non-coding sRNA gene transcribed from the nucleotide sequence DNA shown in the sequence listing.

[0053] In this example, the steps for constructing a recombinant strain of Escherichia coli TOP10 with oxidative stress resistance and / or salt stress resistance are: transforming the constructed pSAncR03 plasmid vector into the strain by heat shock transformation, constructing a recombinant expression The strain E. coli TOP (pSAncR03) is a recombinant Escherichia coli Top10 strain with high resistance to oxidative stress and / or salt stress.

[0054] Verification experiment...

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Abstract

The present invention discovers and confirms the core sequence fragment from the non-coding sRNA gene of Pseudomonas stutzeri for the first time, and combines the Hfq transcription regulator of the non-coding RNA of Escherichia coli to artificially synthesize a new sRNA. Experiments have confirmed that the artificial sequence has the function of endowing host cells with anti-oxidation and high-salt stress tolerance.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a gene capable of improving host cell adversity stress resistance, and specifically relates to an artificially synthesized sRNA with anti-oxidation and high-salt stress tolerance, and the invention also relates to the method of artificially synthesizing the sRNA use. Background technique [0002] sRNA was first discovered in E. coli forty years ago, but its coding gene and function have not been determined. Recent studies have shown that sRNA controls different cellular activities in bacteria, such as acid tolerance, glucose metabolism and outer membrane stress response, etc. The transcriptional activation of OxyS sRNA can protect cells from oxidative stress, while the activation of DsrA sRNA can protect cells from low temperature damage. [0003] Hfq is a post-transcriptional regulator. In recent years, more and more studies have shown that sRNA is assoc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/70C12N1/21
Inventor 燕永亮陆伟战嵛华王劲张维陈明林敏
Owner 北京绿氮生物科技有限公司
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