Extraction method for mangrove plant kandelia candel leaf total protein suitable for two-dimensional electrophoresis

A mangrove plant, two-dimensional electrophoresis technology, applied in the field of mangrove plant protein extraction and separation, can solve the problems of less two-dimensional electrophoresis, no good protein extraction method, etc., and achieves less interfering substances, low protein extraction efficiency, and simple operation. Effect

Active Publication Date: 2016-01-13
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
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Problems solved by technology

At present, there are few reports on the application of two-dimensional electrophoresis in mangrove plants, especially in the study of mangrove plant proteomics, there is no good protein extraction method

Method used

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  • Extraction method for mangrove plant kandelia candel leaf total protein suitable for two-dimensional electrophoresis
  • Extraction method for mangrove plant kandelia candel leaf total protein suitable for two-dimensional electrophoresis
  • Extraction method for mangrove plant kandelia candel leaf total protein suitable for two-dimensional electrophoresis

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Embodiment 1

[0036] At present, the commonly used plant protein extraction methods mainly include TCA-A method and Phe method. Now take mangrove plant kandel (Kandeliaobovata) as material, utilize SDS-PAGE one-way electrophoresis and two-dimensional electrophoresis to compare the extraction effect of TCA-A method, Phe method and the Phe-B method of the present invention, wherein the Phe-B method of the present invention Include the following steps:

[0037] 1. Extraction of candelabra leaf protein by Phe-B method:

[0038] (1), after getting 2g of fresh candelabra leaves and cutting them into pieces, add 10% PVPP (cross-linked polyvinylpyrrolidone) (that is, add 0.1gPVPP per 1g sample) to fully grind with liquid nitrogen in a mortar, and transfer the ground powder to in pre-cooled centrifuge tubes;

[0039] (2) Wash the powder in the centrifuge tube of step (1) with TCA / acetone, add 10ml of pre-cooled acetone containing 10% TCA in mass-volume ratio, vortex for 1min, and shake fully at 4°...

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Abstract

The invention discloses an extraction method for mangrove plant kandelia candel leaf total protein suitable for two-dimensional electrophoresis. A traditional trichloroacetic acid/acetone precipitation method for protein extraction and a traditional phenol extraction method for protein extraction are combined, a cosolvent SDS solution is introduced, the extraction process is optimized, and the Phe-B method suitable for extraction of the mangrove plant kandelia candel leaf total protein is established. The extraction efficiency and quality of the protein are effectively improved, and the technical scheme can be specifically suitable for extraction of the kandelia candel leaf protein in two-dimensional electrophoresis. The method has the advantages of being easy to operate, high in protein extraction efficiency and fewer in interfering substance, and is suitable for materials with difficult protein extraction, low protein extraction efficiency and more interfering substances. The extracted kandelia candel leaf protein completely meets the requirements of the first direction and the second direction of two-dimensional electrophoresis, a high-quality two-dimensional electrophoresis gel map with high resolution, more clear protein points, uniform distribution and the clear background can be obtained, and experiment repeatability and stability are good.

Description

Technical field: [0001] The invention relates to a method for extracting and separating protein from mangrove plants in the field of proteomics research, in particular to a stable and efficient method for extracting total protein from leaves of mangrove plant candelabra which is suitable for two-dimensional electrophoresis. Background technique: [0002] Mangrove is a marine wetland biome composed of evergreen shrubs or trees with mangrove plants as the main body, which grows on tropical and subtropical bays and estuary mudflats. It is an important marine wetland ecosystem. Mangroves play an important role and have huge social and economic benefits in terms of wind and wave protection, embankment protection, bank consolidation, silt consolidation, water purification, and maintenance of coastal wetland biodiversity. At present, the global extreme weather and climate have a direct impact on the survival and development of mangroves. As a common species of mangrove plants, cand...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 费姣王友绍程皓
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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