Detection method of motor protein activity

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

Active Publication Date: 2021-04-06
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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  • Detection method of motor protein activity
  • Detection method of motor protein activity
  • Detection method of motor protein activity

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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 application discloses a method for detecting the activity of motor protein, which comprises the following steps: (1) firstly mix different concentrations of adenosine triphosphate ATP with luciferin / luciferase to obtain the mixture; (2) divide the mixture into two equal parts Add the test sample pool and the control sample pool respectively; (3) inject the buffer solution into the control sample pool, and inject the drive motor into the test sample pool; (4) turn on the detector to detect the product ADP and fluorescein in the test sample pool The fluorescence intensity produced by the / luciferase reaction and the fluorescence intensity in the control sample pool obtain the fluorescence intensity time curve; (5) for the ATP of different standard concentrations, measure the corresponding F ATP , and then generate a calibration curve; (6) make a Mie curve: (7) according to the Mie formula, the fitted Vmax is the active parameter for measuring the hydrolysis of ATP by the driving motor; (8) judge the activity of the driving motor. The detection method of the invention can conveniently and quickly detect the activity of the driving motor protein.

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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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N21/64
CPCG01N21/6428G01N33/6872
Inventor 舒咬根罗永涛
Owner YISHANG TOMORROW TECH
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