Repressor protein, regulatory element group, gene expression regulation system and construction method thereof

A repressor protein and structural domain technology, applied in the field of synthetic biology, can solve problems such as the inability to universally apply repressor engineering, and achieve the effects of improving transcriptional repression ability, reducing the probability of binding, and increasing steric hindrance

Active Publication Date: 2017-11-14
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This excellent effect shows the potential of engineered repressors to achieve network dynamic regulation, however, this network construction method is largely dependent on the specific properties of FapR and cannot be generally applied to engineer repressors

Method used

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  • Repressor protein, regulatory element group, gene expression regulation system and construction method thereof
  • Repressor protein, regulatory element group, gene expression regulation system and construction method thereof
  • Repressor protein, regulatory element group, gene expression regulation system and construction method thereof

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Embodiment

[0124] In the embodiment of the present invention, 9 completely different wild-type repressor proteins and their operating sites known in the prior art are engineered to prepare ultra-sensitive transcriptional control elements with different transcriptional control strengths, and the multimerization structure The effects of adding repressor proteins to domains and / or operator sites to promoter regions are demonstrated. The wild-type repressor protein and its operating site are shown in Table 1.

[0125] Table 1 Wild-type repressor protein and its corresponding operator site sequence

[0126]

[0127]

[0128] 1. Engineering of repressor protein

[0129] The CI multimerization domain (corresponding to the 136-236th amino acid of the CI protein, the DNA sequence encoding the domain is SEQ ID NO: 25) or the CI434 multimerization domain (corresponding to the 70-236th amino acid of the CI434 protein) 210 amino acids, the DNA sequence encoding this domain is SEQ ID NO: 26) f...

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Abstract

By introducing a multimerized structural domain into a repressor protein and optimizing the number of operator loci interacting with the repressor protein in the preferred embodiment, the invention obtains the repressor protein, a regulatory element group and a gene expression regulation system which have ultrasensitive regulation performance.

Description

technical field [0001] The invention belongs to the field of synthetic biology, and relates to an engineered repressor protein, an ultrasensitive regulatory element group, a gene expression regulation system and their construction methods. Background technique [0002] Synthetic biology "programs" cells with the help of standardization of biological components and conceptual abstraction, allowing artificially designed genetic networks to perform various tasks. In recent years, with the rapid development of synthetic biology, the connection between network structure and function has been gradually established, and the predictability of gene circuits and pathways has been greatly improved, making accurate detection of protein nodes, pathways and even networks. and dynamic control becomes possible. [0003] A current research hotspot is the use of synthetic biology to dynamically regulate cellular metabolic pathways in fermentation engineering [1-3]. Artificially constructed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/63
CPCC07K14/47C12N15/63C12N2830/005
Inventor 娄春波侯君然
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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