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Method for separating and detecting antifreeze protein of alpine periglacial plant, and equipment

A technology of antifreeze protein and high mountain, which is applied in the field of protein engineering, can solve the problems of high requirements for instruments, finding, and difficulty in mastering apoplast extraction technology, and achieve the effect of low requirements for instruments

Inactive Publication Date: 2005-06-15
LANZHOU UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the non-universal distribution of antifreeze proteins, it is difficult to find them from certain plants, and the traditional apoplast extraction technology is difficult to master
Low-temperature cycler temperature control and inverted phase contrast microscopy technology have high requirements for instruments, which is difficult for general laboratories

Method used

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  • Method for separating and detecting antifreeze protein of alpine periglacial plant, and equipment
  • Method for separating and detecting antifreeze protein of alpine periglacial plant, and equipment
  • Method for separating and detecting antifreeze protein of alpine periglacial plant, and equipment

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Embodiment Construction

[0020] The present invention will also be further described in detail in conjunction with the accompanying drawings: first, the alpine periglacial plant leaves Rhodiola alpine, alpine bluegrass, alpine ion mustard, and Polygonum spp. are ground with liquid nitrogen, and the powder obtained by grinding is used for Mix the pH 8.0 phosphate buffer and centrifuge at 12,000g for 10 minutes. Take the supernatant, which is the crude extract of soluble protein. Put 45ml of the crude extract into a 50ml small beaker and store it at 4°C for later use. Use high-pressure nitrogen through the pressure reducing valve 2 to flow through the liquid nitrogen along the conduit 3 with a pressure of 0.1 MPa to pre-cool the probe 7. The probe 7 is immersed in the 50 ml small beaker 5 containing the crude protein extract for freezing and ice adsorption, and the rotor is placed inside for magnetic force extraction. Stir to keep the solution homogeneous (see figure 1 ). When the liquid level in the b...

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Abstract

This invention relates to a method for a mountain ice field plant tissue culture and generating cold derivation secretory protein which grinds mountain ice field plant leaves stored in liquid nitrogen to mix the powder evenly with phosphoric acid buffer liquid and to decenter 12000g for a period of time to get the rough extracted solution of soluble protein. An ice crystal adsorber device is used to enrich the anti-frozen protein for two times to be desalinated by a dialysis bay and concentrated, then an ice crystal suppressing observation device is used to carry out recrystallization active test to the quantitative protein.

Description

technical field [0001] The invention belongs to protein engineering technology, and relates to a method and a device for preliminary separation and detection of antifreeze protein of alpine periglacial plants. Background technique [0002] Antifreeze proteins were first discovered in Antarctic fish. They are special proteins that can inhibit the growth of ice crystals. They can inhibit the recrystallization process and lower the freezing point of solutions. So far, antifreeze proteins have been found in fish, insects, fungi and plants, but The homology between these proteins is very low, which proves that this is a newly evolved protein, so the best solution to find is to separate and detect antifreeze proteins from species in low temperature environments. The antifreeze activity of plant antifreeze proteins in rye, ryegrass, and carrots to inhibit recrystallization is superior to that of fish or other sources of antifreeze proteins, and more and more data prove that the mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N33/68
Inventor 安黎哲王建辉
Owner LANZHOU UNIVERSITY