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Method for preparing fluorescent magnetic nanoparticles with streptavidin combination function

A magnetic nanoparticle and nanoparticle technology, applied in the field of nanomaterials, can solve the problems of poor water solubility, low quantum yield, and high viscosity of inorganic light-emitting quantum dots, and achieve selective separation and purification, good water solubility, and small toxic and side effects Effect

Inactive Publication Date: 2011-04-13
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

However, due to their poor water solubility, high viscosity, low quantum yield, and poisonous metal cadmium compounds, inorganic luminescent quantum dots are limited in biomedical and other fields.

Method used

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  • Method for preparing fluorescent magnetic nanoparticles with streptavidin combination function
  • Method for preparing fluorescent magnetic nanoparticles with streptavidin combination function
  • Method for preparing fluorescent magnetic nanoparticles with streptavidin combination function

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Embodiment

[0034] a. Gene cloning

[0035] Get the sequences of the apcA, apcB, cpcS, cpcU, hol, pcyA genes of Synechocystissp.PCC6803 from the National Center for Biotechnology Information (NCBI) database (http: / / www.ncbi.nlm.nih.gov / ) , and refer to the Strep II tag gene sequence in the literature to design primers. The primers and reaction conditions used are listed in Table 1.

[0036] Using Synechocystis sp.PCC6803 genomic DNA as a template, PCR cloning was performed with the primers in the following Table 1 to obtain the Strep II-apcA gene fragment, the phycobilin biosynthetic enzyme hol and pcyA gene fragments, the StrepII-apcB gene fragment, and the chromophore Lyase cpcS and cpcU gene fragments.

[0037] PCR reaction system: double distilled water 18.7μl, containing Mg 2+ 2.5 μl of buffer, 0.5 μl of dNTP, 1 μl of upstream primer, 1 μl of downstream primer, 0.3 μl of Taq enzyme, and 1 μl of Synechocystis sp. PCC 6803 genome.

[0038] Since in the primer design, the Strep II tag...

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Abstract

The invention belongs to the technical field of nano materials and particularly relates to a method for preparing fluorescent magnetic nanoparticles with streptavidin combination function. The method comprises the following specific steps of: respectively connecting a Strep II label gene to N ends of alpha and beta sub-gene apoprotein genes to obtain Strep II-apcA and Strep II-apcB gene segments; embedding the Strep II-apcA gene segment to the downstream of a 6*His gene, and cloning to one expression vector together with a phycobilin biosynthetic enzyme gene; embedding the Strep II-apcB gene segment to the downstream of the 6*his gene, cloning to another vector together with a chromphore lyase gene, simultaneously converting the two expression vectors into escherichia coli, screening engineering bacteria, separating and purifying the engineering bacteria by protein, and oscillating and mixing the purified double-label recombinant APC fluorescent protein and zinc ion modified superparamagnetism silicon shell nanoparticles to obtain the fluorescent magnetic nanoparticles with the streptavidin combination function. The obtained double-label recombinant protein can biologically combine streptavidin without being modified chemically.

Description

technical field [0001] The invention belongs to the technical field of nanometer materials, and in particular relates to a preparation method of streptavidin-binding functional fluorescent magnetic nanoparticles. Background technique [0002] In the 1980s, with the birth and development of microscopic characterization and manipulation technologies such as scanning tunneling microscope (STM) and atomic force microscope (AFM), nanotechnology came into being. Among them, magnetic nanoparticles have attracted more and more attention due to their good superparamagnetism and easy surface functionalization. Surface-modified superparamagnetic nanoparticles can be applied to biological immobilization, separation, modification and detection, so as to extract or measure a series of biologically active compounds, xenobiotic compounds, intracellular components or cells themselves. [0003] In addition, fluorescently labeled probe technology, as a commonly used bioanalysis technology, ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/63C07K14/405C09K11/06H01F1/11
Inventor 陈英杰秦松刘少芳崔玉琳姜鹏陈华新李富超
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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