Method for marking protein polymer by using quantum dots

A technology for labeling proteins and quantum dots, applied in the field of quantum dot-labeled protein polymers, can solve the problems of poor stability of QDs-labeled proteins, and achieve the effects of high binding stability, solving instability and high repeatability

Active Publication Date: 2014-01-08
铜陵汇泽科技信息咨询有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is: in order to study the influence of three-dimensional protein on the assembly characteristics of quantum dots and proteins, and the problem of poor stability of QDs-labeled proteins, in order to solve the above technical problems, the present invention provides a quantum dot-labeled protein aggregation In the object method, a fluorescent protein dimer containing Histag was selected to improve the stability of quantum dots and protein binding

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  • Method for marking protein polymer by using quantum dots
  • Method for marking protein polymer by using quantum dots
  • Method for marking protein polymer by using quantum dots

Examples

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Embodiment 1

[0020] Select CdSe / ZnS quantum dots with an emission wavelength of 565nm, select TIP 1 Protein, the C-terminus is connected to mCherry, and the N-terminus is connected to a Histag to form a TIP 1 -mCherry monomer. Put two TIP 1 -mCherry monomer via dimeric peptide ligand (CGGWRESAI) 2 connected into a stable H-shape, the TIP 1 -mCherry dimer has 2 Histags ( figure 1 ). Will TIP 1 -mCherry monomer and its dimer are coupled and mixed with quantum dots in different proportions through Histag to obtain TIP 1 -mCherry-QD complex and TIP 1 -mCherry dimer-QD complex for FRET assay ( figure 2 ).

[0021] To detect TIP 1 - Stability of mCherry monomer and its dimer binding to QDs, combining QDs with TIP 1 -After mixing mCherry monomer and its dimer, add imidazole to replace the ligand (final concentration of imidazole is 200mM or 400mM). Imidazole can chelate Zn ions, so it can compete with Histag, and it is possible to bind TIP bound to QDs 1 -mCherry monomer and its dim...

Embodiment 2

[0023] Select CdTe / ZnS quantum dots whose emission wavelength is 612nm, and other steps are the same as in Example 1.

Embodiment 3

[0025] The fusion protein used was TIP 1 -GFP fluorescent protein, other steps are the same as in Example 1.

[0026] It can be seen from the above three examples that the present invention can realize the rapid combination of quantum dots and dimer proteins, and has high stability and convenient operation.

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Abstract

The invention discloses a method for marking a protein polymer by using quantum dots and belongs to the field of a nano-biotechnology. The method is characterized by the following steps: selecting a fluorescent protein dimer (TIP1-mCherry dimer) containing a histidine tag sequence (Histag); and mixing protein and the quantum dots according to a certain ratio and coupling to form a protein-quantum dot composite, wherein the structure of the composite is proved to be very stable through imidazole substitution. According to the method, systematic research is performed on the influence of the assembling characteristic of the protein with a three-dimensional structure and the quantum dots, so the combining stability of the protein and the quantum dots is improved and the marked protein polymer can serve as a nanometer fluorescent probe widely applied to a biomarker.

Description

technical field [0001] The invention relates to the field of nanobiology technology, in particular to a method for marking protein polymers with quantum dots. Background technique [0002] Quantum dots (QDs) are zero-dimensional semiconductor nanocrystals, approximately spherical, with a diameter of 1-12nm, and can be dispersed in water or organic solvents to form glue. Compared with traditional organic dyes, QDs have more excellent spectral properties, such as wide and continuous distribution of excitation spectrum, and symmetrical distribution of emission spectrum with narrow width, adjustable color, and high photochemical stability, not easy to photolysis (Marcel BJ , Mario M, Peter G, et al. Science 1998, 281, 2013-2016). These superior properties make QDs widely used in in vivo cell imaging and biological specific labeling. It is and will increasingly become a very important probe tool in molecular cell imaging research. [0003] At present, fat-soluble CdSe / ZnS quant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/13
Inventor 王建浩蒋鹏举邱琳王车礼李静燕郭沛霖李进晨夏江
Owner 铜陵汇泽科技信息咨询有限公司
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