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Method for labeling protein through quantum dots

A technology for labeling proteins and quantum dots, applied in the field of nanobiology, can solve the problems of poor stability of proteins and QDs, achieve high repeatability, simple operation methods, and solve instability effects

Inactive Publication Date: 2013-02-13
CHANGZHOU UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved in the present invention is: in order to overcome the problem of poor stability of existing proteins and QDs and limit their popularization and use in the biological field, in order to solve the above technical problems, the present invention provides a new method for labeling proteins with quantum dots, Utilizing the property of SpeA protein that is prone to dimerization, the resulting dimer produces two Histags, thereby improving the stability of the binding of quantum dots to proteins

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  • Method for labeling protein through quantum dots

Examples

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Embodiment 1

[0018] SpeA protein expression with Histag

[0019] The SpeA gene was purchased from Genscript and expressed in Escherichia coli (E. coli). The gene was cloned into the NdeI and EcoRI sites of the pET28a vector to obtain a high-yield expression plasmid pJX2, which contained the full-length SPEA protein and the N-terminal Histag. Plasmid pJX2 was introduced into E. coli by electroporation. After the recombinant gene is transformed into the E. coli strain, it is induced to express the target protein in the host bacteria through temperature control, and the expression sample is subjected to SDS-PAGE to detect the expressed protein. SpeA will form a dimeric structure in the solution ( figure 2 ).

[0020] After mixing SpeA and CdSe / Zns quantum dots at a molar ratio of 8:1, shake for 30 minutes, and the SpeA dimer is combined with the quantum dots through histidine to obtain a SpeA-QD complex ( figure 1 ).

[0021] To test the stability of SpeA binding to QDs, after mixing QD...

Embodiment 2

[0023] The quantum dots used are CdTe / ZnS, the molar ratio of protein to quantum dots is 4:1, other steps are the same as in Example 1.

Embodiment 3

[0025] The quantum dots used are CdSe / ZnSe, the molar ratio of protein to quantum dots is 16:1, and 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 SpeA protein dimer, and has high stability and convenient operation.

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Abstract

The invention discloses a method for labeling protein through quantum dots and belongs to the field of nano-biotechnology. The method is characterized by expressing a Histag on protein, obtaining a protein dimer containing two Histags through protein dimerization, mixing the protein with the quantum dots according to a certain ratio, and oscillating to enable the two Histags to combine with the quantum dots simultaneously so as to improve the stability of combination. The method is simple in operation, and the quantum dots can be applied to biomarker widely as a nano fluorescent probe.

Description

Technical field [0001] The present invention involves a new method of quantum dot marking protein in the field of nano -biotechnology. Background technique [0002] Quantum dot (QDS) is a zero -dimensional semiconductor nano -crystal, which is similar to spherical and diameter 1 ~ 12nm. It can be scattered in water or organic solvents to form glue.Compared with traditional organic dyes, QDS has a better spectral properties, such as stimulating the spectral wide and continuous distribution, and the launch spectrum is symmetrically distributed and narrowed, the color can be adjusted, and the optical chemical stability is high., Mario M, Peter G, et al. Science 1998, 281, 2013-2016).These superior properties have widely used QDS cell imaging and biological specific markers in the body. It is increasingly becoming a very important probe tool in molecular cell imaging research. [0003] At present, the most widely used in biomarkers is the fat -soluble CDSE / ZNS quantum dot synthesized...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/13C07K14/315
Inventor 王建浩王车礼蒋鹏举邱琳夏江
Owner CHANGZHOU UNIV
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