Extraction and dielectrophoresis analytical method for total protein of cattail cauloid
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A technology of two-dimensional electrophoresis and analysis method, which is applied in the biological field to achieve the effects of good repeatability, high extraction rate and clear map.
Active Publication Date: 2011-11-16
常熟紫金知识产权服务有限公司
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The main nutrients and medicinal substances in cattail are protein compounds, but the plants also contain more secondary metabolites (such as: pigments, phenols, quinones, etc.), which have not been targeted at in the prior art. Quality Analysis Method of Protein Components in Pucai
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Embodiment 1
[0045] The indoor hydroponic cattail pseudostem total protein extraction and two-dimensional electrophoresis analysis method specifically includes the following steps:
[0046] (1) Collect indoor hydroponic cattail pseudostems, wash them with ultrapure water, absorb excess water with filter paper, put them into liquid nitrogen for quick freezing, and move them to a -80°C refrigerator for future use;
[0047] (2) take by weighing the cattail pseudostem of 5g step (1) preservation and put into the mortar of ice bath rapidly, add liquid nitrogen and grind into fine powder, add 0.5g polyvinylpyrrolidone in the grinding process, turn the fine powder after grinding Put it into a 50ml centrifuge tube pre-cooled at -20°C, add 3 times the volume of -20°C pre-cooled trichloroacetic acid (TCA) / acetone solution, mix well, put it in a -20°C refrigerator for 2 hours, and set aside; The trichloroacetic acid (TCA) / acetone solution described therein consists of the following substances: trichl...
Embodiment 2
[0069] The extraction and two-dimensional electrophoresis analysis method of total protein in the cattail pseudostem planted along the lake in Huai'an City, Jiangsu Province specifically includes the following steps:
[0070] (1) Collect pseudostems of cattails planted along the lake in Huai'an City, Jiangsu Province, wash them with ultrapure water, absorb excess water with filter paper, put them into liquid nitrogen for quick freezing, and move them to a -80°C refrigerator for future use;
[0071] (2) take by weighing the cattail pseudostem of 5g step (1) preservation and put into the mortar of ice bath rapidly, add liquid nitrogen and grind into fine powder, add 0.5g polyvinylpyrrolidone in the grinding process, turn the fine powder after grinding Put it into a 50ml centrifuge tube pre-cooled at -20°C, add 3 times the volume of -20°C pre-cooled trichloroacetic acid (TCA) / acetone solution, mix well, put it in a -20°C refrigerator for 2 hours, and set aside; The trichloroaceti...
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Abstract
The invention discloses an extraction and dielectrophoresis analytical method for total protein of cattail cauloid, comprising the following steps of: firstly, efficiently extracting protein from the cattail cauloid by screening through many experiments; under a preferred condition, separating the protein components from the secondary metabolite (such as pigments, phenols, quinones and the like) components by a dielectrophoresis method; and performing qualitative analysis by a silver nitrate staining method under a preferred condition, and / or performing qualitative and quantitative analysis on the separated protein by a mass spectrometer. Shown by experimental results, more protein spots are obtained by separation through the dielectrophoresis analytical method, and detected by PDQuest image analysis software, the number of the protein spots is more than 1000, and the spectrum is clear without transverse and longitudinal stripes; and the method has good repeatability and stability, can be used for conveniently identifying the authenticity of the pharmaceutical / food resource plant cattail, and provides a quality control standard and scientific base for in-depth study, scientific planting, culture, development and utilization of cattail.
Description
technical field [0001] The invention relates to a plant quality analysis and identification method, in particular to a method for extracting and two-dimensional electrophoresis analysis of total protein in cattail pseudostems, and belongs to the field of biotechnology. Background technique [0002] cat-tail( Typha latifolia L.), also known as cattail cattail, cattail cattail, water candle, is a perennial swamp herb of the genus Typhaceae, distributed in swamps and freshwater lakes all over the world. In some areas of our country, the young and tender part of the vegetative body is used as a vegetable, which has a unique flavor and rich nutrition, and it also has medicinal value. Therefore, it is a health-care vegetable that is rich in nutrition and has certain medicinal functions. The main nutrients and medicinal substances in cattail are protein compounds, but the plants also contain more secondary metabolites (such as: pigments, phenols, quinones, etc.), which have no...
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