Method for marking enveloped virus nucleocapsid by using quantum dot

A technology of quantum dots and nucleocapsids is applied in the field of quantum dots marking enveloped virus nucleocapsids, which can solve the problems of interfering with virus identification virus infection activity, and achieve the effect of maintaining infection activity and a mild method.

Inactive Publication Date: 2014-02-05
WUHAN UNIV
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Problems solved by technology

However, at present, quantum dots can only be used to mark the envelope of enveloped viruses, which may interfere with the recognition of the virus on cell receptors and affect the infection activity of the virus. Therefore, using quantum dots to mark the nucleocapsid inside the enveloped virus is a a better option

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  • Method for marking enveloped virus nucleocapsid by using quantum dot
  • Method for marking enveloped virus nucleocapsid by using quantum dot
  • Method for marking enveloped virus nucleocapsid by using quantum dot

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Embodiment 1

[0023] Example 1 Quantum dot-labeled recombinant baculovirus nucleocapsid

[0024] The method of the present invention will be described in detail below by taking the quantum dot labeling of baculovirus nucleocapsid as an example.

[0025] 1. Method

[0026] 1. Construction of recombinant baculovirus plasmid: using the baculovirus expression system, simultaneously insert the VP39AP sequence (the baculovirus capsid protein VP39 sequence fused with the biotin receptor peptide AP sequence) and the ability to The BirA enzyme sequence that catalyzes AP biotinylation, thereby obtaining a plasmid with the entire genome of the recombinant baculovirus.

[0027] 2. Obtain the purified recombinant baculovirus RBV: transfect the recombinant plasmid into the host Sf9 cells with a transfection reagent, collect the cell supernatant when all the cells have lesions after 72 hours, and pass it at 5000 rpm for 15 min at 4°C Centrifuge to obtain the first generation of recombinant baculovirus...

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Abstract

The invention relates to a method for marking enveloped virus nucleocapsid by using a quantum dot. The method is characterized in that a capsid protein sequence blended with a biotin receptor peptide sequence and a catalytic receptor peptide biotinylation enzyme sequence are simultaneously inserted in a virus whole-genome so as to obtain a recombinant virus plasmid, the biotinylation of the nucleocapsid is realized by virtue of the reproduction process of the virus plasmid in a host cell, and then the quantum dot coupled with streptavidin is used for realizing the marking of the enveloped virus nucleocapsid. The method does not needs the violent chemical reaction transformation and the marking of a virus and is beneficial to the maintenance of infection activity of the virus self; and the marking of the nucleocapsid in the enveloped virus does not affect the surface of the enveloped virus and the indentation of the surface. The method can be applied to the virus infection mechanism study and the virus-related disease treatment research.

Description

[0001] technical field [0002] The invention belongs to the fields of chemistry and biomedicine, and relates to a method for marking enveloped virus nucleocapsids with quantum dots. Background technique [0003] The research on the mechanism of virus infecting cells is crucial to the prevention and treatment of diseases caused by viruses, and the fluorescent labeling of live viruses can provide an important basis for studying the mechanism of virus infection. Traditional fluorescent markers often have disadvantages such as easy photobleaching, while semiconductor fluorescent quantum dots have unique optical properties, such as high fluorescence quantum yield, wide excitation spectrum, and narrow emission spectrum. way of the virus. Enveloped viruses widely exist in nature and are closely related to human health, so they have received great attention in related research. However, at present, quantum dots can only be used to mark the envelope of enveloped viruses, which may...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12Q1/70C12R1/93
Inventor 庞代文文莉林毅张志凌王汉中
Owner WUHAN UNIV
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