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Kinesin activity detection method

A technology for driving motor and protein activity, which is used in biological testing, measuring devices, material inspection products, etc., to achieve the effect of convenient and fast detection

Active Publication Date: 2018-11-13
YISHANG TOMORROW TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the prior art, there is no convenient and quick method for detecting the activity of motor protein

Method used

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Embodiment Construction

[0046] The application will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain related inventions, rather than to limit the invention. It should also be noted that, for ease of description, only parts related to the invention are shown in the drawings.

[0047] It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present application will be described in detail below with reference to the accompanying drawings and embodiments.

[0048] Detection principle of the present invention:

[0049] A normal driving motor can travel more than 800nm ​​per second under the saturated ATP concentration, that is, consume more than 100 ATP per second, and the ATP concentration in the solution can be quantified by the luciferin / luciferase...

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Abstract

The invention discloses a kinesin activity detection method. The method comprises the following steps: (1) ATP (adenosine triphosphate) with different concentrations is mixed with fluorescein / luciferase, and mixtures are obtained; (2) the mixtures are equally divided into two parts and added to a test sample pool and a control sample pool respectively; (3) a buffer solution is injected into the control sample pool, and kinesin is injected into the test sample pool; (4) a detector is started for detecting fluorescence intensity obtained by a reaction of the product ADP in the test sample pool with fluorescein / luciferase and fluorescence intensity of the control sample pool, and a fluorescence intensity-duration curve is obtained; (5) corresponding FATP is measured for ATP with different standard concentrations, and a calibration curve is generated; (6) a Michaelis-Menten curve is drawn; (7) according to a Michaelis-Menten formula, Vmax obtained by fitting is an active parameter for measuring ATP hydrolysis of kinesin; (8) judging activity of kinesin. The activity of kinesin can be detected conveniently and quickly with the detection method.

Description

technical field [0001] The present application relates to a detection method, in particular to a detection method for motor protein activity. Background technique [0002] Kinesins are important intracellular transport machinery and belong to translational molecular motors. It has two main features: one is persistence, at least one of the two heads of the motor remains adsorbed to the microtubules when walking alternately, so the motor can walk along the microtubules for a long distance without derailment; the other feature is The mechanical and chemical processes of the motor are tightly coupled, that is, the motor consumes one adenosine triphosphate (ATP) per step forward. [0003] In the prior art, there is no convenient and quick method for detecting the activity of kinesin. Contents of the invention [0004] In view of the above defects or deficiencies in the prior art, it is desired to provide a method for detecting the activity of the kinesin protein conveniently ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/64
CPCG01N21/6428G01N33/6872
Inventor 舒咬根罗永涛
Owner YISHANG TOMORROW TECH
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