Plant plasma membrane protein redissolution method and application in two-dimensional fluorescent difference gel electrophoresis thereof

Abstract

The invention discloses a plant plasma membrane protein redissolution method and application in two-dimensional fluorescent difference gel electrophoresis thereof. The plant plasma membrane protein redissolution method comprises the following steps: plant plasma membrane protein is added to a lysis solution, ultrasonic processing is conducted in an ice bath, then the plant plasma membrane protein is dissolved by eddy mixing, and at last supernatant fluid is collected in a centrifugal mode, namely a plant plasma membrane protein solution is obtained. The lysis solution is obtained by that N - dodecyl - beta - D - malt sugar is added to a fluorescent difference gel electrophoresis (DIGE) lysis buffer solution, and the concentration of the DIGE lysis buffer solution is set to be 0.5-2 g / 100 ML. The plant plasma membrane protein is low in abundance degree and high in hydrophobic property, certain difficulty exists in the extraction and the redissolution, and thus great challenge is brought to the research of plasma membrane proteomics. According to the plant plasma membrane protein redissolution method, the extraction efficiency of the plant plasma membrane protein is increased by nearly two times, the electrophoresis result of good repeatability and high resolution ration can be obtained after the 2-DDIGE is conducted, and more protein points are displayed compared with an existing method.

Description

technical field [0001] The invention relates to a method for redissolving plant plasma membrane proteins and its application in two-way fluorescent differential gel electrophoresis. Background technique [0002] The plant cell plasma membrane is the barrier between the cell and the external environment. It is also a key part of plant cells for material exchange, energy conversion, and information transmission. important role in the process. The main executors of plasma membrane function are plasma membrane proteins. However, plant plasma membrane proteins have low abundance and strong hydrophobicity, and most plasma membrane proteins have post-translational modifications such as glycosylation and phosphorylation, resulting in microscopic inhomogeneity of membrane proteins. Therefore, how to separate, purify and identify cell membrane proteins is a difficult point in the study of plant plasma membranes. [0003] The separation and identification of plant cell plasma membran...

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Problems solved by technology

[0004] At present, there have been some experiments on the analysis of plant plasma membrane proteins by two-dimensional electrophoresis technology, but there are many problems: the extraction efficiency of plant plasma membrane proteins is low, and the protein quantity required by two-dimensional electrophoresis technology is large; plasma membrane proteins are hydrophobic Strong, insoluble, and insoluble plasma membrane proteins are difficult to enter the gel, which affects the later separation and identification

[0006] The research on plant plasma membrane proteins mostly focuses on sweet soil plants (such as rice, soybean and other model plants), while halophytes are an important group of plants, and the plasma membrane plays an important role in their salt adaptation process, but about salt There are few studies on plasma membrane proteins of raw plants

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