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Oxidation and high salt stress resistant artificially synthesized sRNA and application thereof

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, and no host adversity stress, etc., and achieve a high survival rate. , the effect of improving oxidative stress resistance and salt stress resistance

Active Publication Date: 2015-05-06
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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  • Oxidation and high salt stress resistant artificially synthesized sRNA and application thereof
  • Oxidation and high salt stress resistant artificially synthesized sRNA and application thereof
  • Oxidation and high salt stress resistant artificially synthesized sRNA and application thereof

Examples

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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 shown that the loss of sRNA03 will lead to the reduction of the antioxidant and salt tolerance of cells, and the transfer of this sRNA into Escherichia coli will also significantly enhance its ability to survive in peroxidative and high-salt environments, which regulates the physiological process of cell stress resistance has been confirmed. In order to construct an 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...

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

A core sequence fragment from a pseudomonas stutzeri non-coding sRNA gene is discovered and determined for the first time by the invention, and the core sequence fragment is combined with the transcriptional regulon Hfq of an escherichia coli non-coding RNA to synthesize a novel sRNA artificially. Experiments show that the artificially synthesized sequence endows a host cell with an oxidation and high salt stress resisting function.

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 Applications(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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