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Extraction method of plant total protein and special extract for the same

A technology of plant total protein and extraction method, which is applied to the preparation of plant peptides, test samples, peptides, etc., and can solve the problems of difficult protein solubility and no way to remove salt ions

Inactive Publication Date: 2007-02-07
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is mainly used in animals and some model plants; in addition, the obvious defect of this method is that the protein precipitated by TCA is difficult to dissolve, and there is no way to remove the salt ions in the sample

Method used

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  • Extraction method of plant total protein and special extract for the same
  • Extraction method of plant total protein and special extract for the same
  • Extraction method of plant total protein and special extract for the same

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Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1, TCA method, E-TCA method, phenol extraction method and BPP method of the present invention extract the one-dimensional electrophoresis comparison of the total protein extraction effect of Salicorniaeuropaea

[0038] At present, the commonly used plant protein extraction methods mainly include TCA, E-TCA and phenol extraction method. Now, the euhalophyte Salicornia europaea (Salicornia europaea) treated with different concentrations of NaCl is used as the material, and the TCA is compared by SDS-PAGE one-dimensional electrophoresis. The extraction effect of method, E-TCA method, phenol extraction method and BPP method of the present invention, concrete method comprises the following steps:

[0039] 1. Salicornia cultivation and salt treatment

[0040] Salicornia has significant salt-accumulating characteristics, and it needs to grow under certain salinity conditions to achieve the best growth (Davy A J, Bishop G F and Costa C B.Salicornia L.(Salicornia pusi...

Embodiment 2

[0055] Example 2, SDS-PAGE one-dimensional electrophoresis detection of different tissue protein extraction effects of different plants using BPP method

[0056] Now use the BPP method of the present invention to extract the total protein of fourteen kinds of plant different tissues, including four kinds of woody plants (pine (Pinus bungeana Zucc.), Canadian poplar (Populus canadensisMoench.), hickory ( Carya illinoinensis K.) and Peach (Prunus persica Rehd.), carried out SDS-PAGE detection to the extraction effect, and the detection results were as follows figure 2 Shown (swimming lanes 1-3 are respectively the total protein extracted from the needle leaf (swimming lane 1), branch phloem (swimming lane 2) and xylem (swimming lane 3) of pine (Pinusbungeana Zucc.); Swimming lanes 4-6 are respectively from Canada Total protein extracted from leaves (lane 4), phloem (lane 5) and xylem (lane 6) of poplar (Populus canadensis Moench.); ), peach (Prunus persica Rehd., lane 8), plan...

Embodiment 3

[0057] Embodiment 3, extract the productive rate of different tissue protein of different plants with BPP method

[0058] The BPP method was used to extract the total protein of sixteen kinds of common plants (see Table 1 for the plant name) in different parts and different tissues, and the protein content was determined, and the protein extraction rate was calculated. rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. AnalBiochem, 1976, 72: 248-254.), using a UV spectrophotometer (Shimadzu UV-160, Kyoto, Japan), and then measuring OD 595 The protein extraction yield was calculated, and bovine serum albumin (BSA) was used as the standard for calibration. The detection results of protein yield are shown in Table 1. BPP method can be used to extract an average of 2.0 mg of protein per gram of fresh plant material. The yield of protein extracted from leaves is higher than that of other tissues. The ...

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Abstract

the invention discloses an extracting method of total plant protein and specific extracting liquid, which comprises the following parts with pH value at 7.5-8.5: 80-120mM EDTA, 80-120mM trimethyl aminomethane, 40-60mM sodium tetraborate and 40-60mM vitamin C, 0.8-1.2% polyvinyl polypyrrolidon,0.8-1.2% Triton-100, 1.5-2.5% 2-mercaptoethanol and 28-32% saccharose solution.

Description

technical field [0001] The invention relates to a method for extracting total plant protein and a special extracting solution thereof in the field of proteomics research. Background technique [0002] In proteomics research, currently the most effective method is to separate proteins by two-dimensional electrophoresis (2-DE), and then identify the separated proteins by mass spectrometry (Grg A, Weiss W, Dunn M J. Current two-dimensional electrophoresis technology for proteomics. Proteomics, 2004, 4: 3665-3685.). Since various protein extraction methods for two-dimensional electrophoresis often vary greatly due to different plant species and tissue parts, extraction of high-quality proteins has become one of the most challenging tasks in two-dimensional electrophoresis and later mass spectrometry identification. one. Plant tissues usually contain a large amount of recalcitrant substances, such as polysaccharides, esters and polyphenols, which will affect the effect of two-...

Claims

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Application Information

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IPC IPC(8): C07K2/00C07K4/10G01N1/34
Inventor 李银心王旭初
Owner INST OF BOTANY CHINESE ACAD OF SCI
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