Drug screening model taking histidine kinase AgrC as target spot

A histidine kinase and drug technology, applied in the field of biomedicine, to achieve the effects of convenient operation, strong specificity and clear action site

Active Publication Date: 2017-02-22
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it has not been reported that a drug screening model based on AgrC proteoliposomes has been successfully screened for a drug monomer compound that has a strong inhibitory effect on the kinase activity of the AgrC protein.

Method used

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  • Drug screening model taking histidine kinase AgrC as target spot
  • Drug screening model taking histidine kinase AgrC as target spot
  • Drug screening model taking histidine kinase AgrC as target spot

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Embodiment Construction

[0034] The technical solutions of the present invention will be further described below in conjunction with specific examples, but the present invention is not limited by the content of the examples in any form. Unless otherwise specified, the test methods described in the examples are conventional methods; unless otherwise specified, the reagents and biological materials can be obtained from commercial sources.

[0035] Escherichia coli strains were purchased from Novagen, Escherichia coli strains BL21(DE3), Tuner(DE3), C41(DE3), C43(DE3), BL21(DE3)-pLysS and BL21-CodonPlus(DE3)-RIL were all acceptable. For the expression of AgrC protein, preferably C43(DE3). The purpose of choosing six Escherichia coli as hosts to express AgrC protein is to investigate which strain has the highest expression level of AgrC protein. After screening, the expression level of AgrC protein in Escherichia coli strain C43 (DE3) was the highest, so this strain was used as a carrier for protein expre...

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Abstract

The invention relates to a drug screening model taking histidine kinase AgrC as a target spot, belonging to the field of biological medicines. A construction method of the drug screening model comprises the following steps: (1) extracting and purifying AgrC protein; (2) determining the concentration of AgrC protein; (3) preparing lipidosome by taking DOPC, DPPA and CHOL as raw materials; (4) inserting the extracted and purified AgrC protein into the lipidosome by adopting a surfactant-mediating method, thus forming protein lipidosome; and (5) constructing the drug screening model taking the histidine kinase AgrC as the target spot. The constructed drug screening model is suitable for screening drug monomeric compounds having a strong inhibiting effect for histidine kinase AgrC, and the screened compounds are definite in acting sites; the drug screening model has the advantages of high flux, strong specificity and operation convenience and rapidness when used for screening the drug monomeric compounds.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a drug screening model whose target is histidine kinase AgrC. Background technique [0002] Accessory gene regulator (agr) is the most important virulence factor regulation system of staphylococcus, which is closely related to the production of various virulence factors and the formation of biofilm. The agr system consists of the signal receptor AgrC and the response regulator AgrA, and belongs to a typical two-component signal transduction system (two component signal transduction system, TCST). Histidine protein kinase AgrC, as the signal receptor of TCST, can be activated by homologous signal molecules in the external environment, undergoes self-transfer phosphorylation, and the phosphorylated AgrC can phosphorylate AgrA, and the activated AgrA can interact with downstream promoters. The repeat sequence between RNAII and RNAIII specifically binds, thereby regulating the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
CPCC12Q1/48
Inventor 权春善张丽影陈莹刘爽熊文范圣第
Owner DALIAN NATIONALITIES UNIVERSITY
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