Method of determining bacillus thuringiensis toxic protein CrylAc

A technology of Bacillus aureus and toxic protein, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of high sensitivity, broad application prospects, and low detection limit

Inactive Publication Date: 2013-06-26
GUILIN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, electrochemiluminescence immunosensors detect transgenic plant proteins, especially based on nucleocapsid Fe 3 o 4 Au nanoparticles sensor for detection of transgenic Bt toxic protein Cry1Ac has not been reported

Method used

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  • Method of determining bacillus thuringiensis toxic protein CrylAc
  • Method of determining bacillus thuringiensis toxic protein CrylAc
  • Method of determining bacillus thuringiensis toxic protein CrylAc

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0025] (1) Fabrication of magnetron glassy carbon electrode (GCE):

[0026] Cut a 3 cm iron rod (diameter 2.55 mm) and a 2 cm glass tube (diameter 3 mm), grind and clean both ends; put glassy carbon into one end of the glass tube, then insert the iron rod into the glass tube and stick it to the glass tube The carbon and iron rods were fixed with molten solid paraffin around the iron rod, and the electrode surface was polished with alumina powder with a particle size of 1.0, 0.3 and 0.05 μm on the suede, and cleaned for use.

[0027] (2) Synthesis of Fe by chemical co-precipitation method 3 o 4 Core:

[0028] in N 2Under protection, 15 mL of NaOH solution with a molar concentration of 2 mol / L was added to 2 g FeSO at 50 °C under vigorous stirring. 4 ·7H 2 O with 0.58 g FeCl 3 in the mixture; keep the pH at 10 during the experiment; continue heating to 80 °C for aging for 1 hour; separate the black precipitate with an external magnet, wash it repeatedly with twice distille...

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Abstract

The invention discloses a method of determining bacillus thuringiensis toxic protein CrylAc. The method comprises the steps that: Fe3O4@Au nana-particles combining with a transgenic Bt toxic protein CrylAc primary antibody is modified onto the surface of a magnetic control glassy carbon electrode, and specific immune reaction is carried out on the magnetic control glassy carbon electrode with an antigen and a GOD labeled secondary antibody to form a compound with a sandwich structure; an electrogenerated chemiluminescence immunosensor is developed to be used for detecting the concentration of transgenic Bt toxic protein CrylAc; glucose is catalyzed and oxidized by using GOD to generate H2O2; voltage is applied to a working electrode to excite luminal to be oxidized; a luminal oxide and the H2O2 react to generate an electrochemiluminescence signal; an electrochemiluminescence response is strengthened along with the increase of the concentration of the GOD; and the electrochemiluminescence signal has a linear relation with the concentration of the transgenic Bt toxic protein CrylAc within a range of 0-6ng / mL. The method has high sensitivity and low detection limit, is widely applied and supplies a wide application prospect of reliably and ultra-sensitively detecting the toxic protein of a transgenic plant.

Description

technical field [0001] The invention relates to a method for measuring the toxin protein Cry1Ac of Bacillus thuringiensis, especially based on Fe 3 o 4 Preparation of electrochemiluminescence immunosensor based on Au nanoparticles and method for determination of Bacillus thuringiensis toxic protein Cry1Ac. Background technique [0002] Nanoparticles have large specific surface area, high surface activity, many surface active centers, and high catalytic efficiency, so they can increase the stability and adsorption capacity of biologically active substances. Fe 3 o 4 Nanoparticles have the characteristics of superparamagnetism, low toxicity, and simple preparation process. However, Fe alone 3 o 4 Nanoparticles are prone to oxidation and agglomeration during storage. Coating a layer of inorganic film on the surface of nanoparticles can overcome the above disadvantages. Fe 3 o 4 The surface of nanoparticles is coated with nano-Au to form a core-shell structure, which c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/76
Inventor 李建平徐倩魏小平
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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