Acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants

A technology of peroxidase and polyacrylamide, applied in the field of electrophoresis, can solve the problems of incomplete zymogram information, unsatisfactory separation effect of magnoliaceae plant peroxidase, etc.

Inactive Publication Date: 2018-11-09
HUBEI UNIV FOR NATITIES
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  • Abstract
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  • Application Information

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Problems solved by technology

[0003] In summary, the problems in the prior art are: The existing technology is not ideal for the separation of magnolia plant peroxidase, and the obtained zymogram information is incomplete, which cannot completely reflect the whole picture of peroxidase site information

Method used

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  • Acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants
  • Acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants
  • Acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants

Examples

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Embodiment 1

[0030] Embodiment 1: Taking Magnoliaceae Magnolia officinalis as an example

[0031] (1) Extraction of Magnolia officinalis leaf peroxidase

[0032] A. Accurately weigh 0.5g of magnolia bark leaves washed, dried and chopped on an analytical balance, add 2.5mL of 50mmol / L pH6.0 acetate buffer extract, grind on ice until homogenized, and then transfer to 5mL centrifuge The tube was centrifuged at 10000 g for 10 min at 4°C, and the supernatant was collected.

[0033] B. Take 0.5mL of the above supernatant in a 2mL test tube, add 0.05mL of sample buffer into it, mix well, centrifuge at 10000g at 4°C for 10min, collect the supernatant, and store it in a refrigerator at 4°C for later use.

[0034] (2) Preparation of electrophoresis reagents

[0035]A. Acrylamide stock solution: Weigh 30g Acr and 1.5g Bis, add water to dissolve, dilute to 100mL, and store in a refrigerator at 4°C in the dark.

[0036] B. 0.5% FeSO 4 : Weigh 0.5g FeSO 4 Add to a small amount of water that has bee...

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Abstract

The invention discloses an acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants, belonging to the technical field of electrophoresis methods. The acidic polyacrylamide electrophoresis separation method for peroxidase in Magnoliaceae plants comprises the following steps: pretreatment and extraction of peroxidase in the leaves of Magnoliaceae plants; unidirectional rapid separation of the peroxidase in the leaves of Magnoliaceae plants; and rapid color development via a benzidine method, and after completion of color development, rapid scanning of dyed gel by selecting a transmission cope mode in an EPSON scanner. According to the invention, acidic polyacrylamide electrophoresis is adopted for separation of the peroxidase in the Magnoliaceae plants, and overcomes the disadvantages of low resolution, small band numbers and the like in traditional polyacrylamide electrophoresis separation of peroxidase. The acidic polyacrylamide electrophoresis separation method of the invention has high application value in the identification, variety breeding and genetic relationship research of Magnoliaceae plants.

Description

technical field [0001] The invention belongs to the technical field of electrophoresis, and in particular relates to an acidic polyacrylamide electrophoresis separation method for magnolia plant peroxidase. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: Plants of Magnoliaceae (Magnoliaceae) are mainly distributed in tropical and subtropical regions of Asia and America. Plants in this family include 2-17 genera and 240-300 species. They are mainly distributed in the southeast to southwest, and gradually decrease in the northeast and northwest. Magnoliaceae plants are important ornamental flowers and trees, traditional Chinese medicine and flavor raw materials in my country, and are widely used in urban landscape design and economic crop cultivation. Therefore, it is of great significance to establish a rapid variety identification method. At present, in the study of classification and identification of Magno...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/44726
Inventor 肖强赵广华周秀梅洪健
Owner HUBEI UNIV FOR NATITIES
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