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A method for screening rev polypeptide small molecule antagonists based on graphene oxide

A graphene and small molecule technology, applied in the field of drug screening, can solve the problems of small polarization change signal range, irregular folding of polypeptide chain, conformational change, etc., and achieve the effect of wide recognition signal range and simple operation

Active Publication Date: 2020-08-25
SHAANXI NORMAL UNIV
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Problems solved by technology

[0004] At present, the screening of Rev polypeptide small molecule antagonists mainly uses fluorescently labeled Rev polypeptide as an indicator. The range of polarization change signals generated by this traditional method is small and the sensitivity is not high. Moreover, fluorescently labeling Rev polypeptide may make the polypeptide chain produce irregular folding or conformational changes

Method used

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  • A method for screening rev polypeptide small molecule antagonists based on graphene oxide
  • A method for screening rev polypeptide small molecule antagonists based on graphene oxide
  • A method for screening rev polypeptide small molecule antagonists based on graphene oxide

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Embodiment 1

[0020] Add proflavin and the HIV-1 RRE RNA fragment (nucleotide sequence: 5'-UG GUCUGGGCGCAGCGCAAGCUGACGGUACAGGCC-3') containing the IIB stem-loop portion to 100 μL of 20 mmol / L pH=7.0 ammonium acetate buffer at 37 Incubate at ℃ for 5 minutes, then add the drug to be tested, continue to incubate at 37℃ for 5 minutes, then add graphene oxide, and mix well. The concentration of some HIV-I RRE RNA fragments is 100nmol / L, the concentration of the drug to be tested is 1 μmol / L, and the concentration of graphene oxide is 8.9 μg / mL, and the LS55 fluorescence spectrophotometer (excitation and emission wavelengths are respectively 455nm and 505nm, the incident slit and the exit slit are both 10nm, and the interval is 1 second) to test the fluorescence polarization degree of the system before and after adding the drug to be tested. If the drug to be tested has an antagonistic effect, the degree of fluorescence polarization is significantly reduced. The screening of Rev polypeptide small...

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Abstract

The invention discloses a graphene oxide based method for screening a Rev polypeptide micro-molecule antagonist. According to the method, proflavine is taken as a fluorescence polarization indicator,graphene oxide is introduced into an action system, the change value of a fluorescence polarization signal indicating whether a Rev polypeptide micro-molecule antagonist exists or not is increased, and the graphene oxide based method for screening a Rev polypeptide micro-molecule antagonist is established. The method has the advantages that the operation is simple, the indicator is not a fluorescence labeled polypeptide, the macromolecular conformation deformation caused by labeling is avoided, at the same time, compared with a conventional fluorescence polarization method, the range of signals on recognizing a Rev polypeptide micro-molecule antagonist is wider, the sensitivity is higher, and the high throughput screening of a Rev polypeptide micro-molecule antagonist is realized.

Description

technical field [0001] The invention belongs to the technical field of drug screening, and in particular relates to a method for screening Rev polypeptide small molecule antagonists based on graphene oxide. Background technique [0002] AIDS is also known as Acquired Immune Syndrome (AIDS), and there is currently no effective cure for it. The causative agent of AIDS is human immunodeficiency virus (HIV), which is a lentivirus that infects cells of the human immune system. Clinically used drugs for the treatment of HIV-I are mainly protease inhibitors and reverse transcriptase inhibitors. These two types of drugs are prone to variability, drug resistance and toxicity. Therefore, exploring new mechanisms of action is not easy to produce drug resistance Drugs have become a hot spot in the research of anti-AIDS drugs. The HIV-I transcription process is mainly regulated by some transcription factors, such as regulator of virion protein expression (Rev) and so on. Rev can affec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6445
Inventor 张志琪亓亮张丹范瑶瑶
Owner SHAANXI NORMAL UNIV
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