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Method for extracting and purifying wild rice stem total proteins

A purification method and a technology of protein extraction solution, which are applied in the field of extraction and purification of total protein of Zizania zizania plants, can solve the problems that the extraction and purification method has not been reported, and achieve the effect of less impurities, low nucleic acid content and high protein content

Inactive Publication Date: 2012-03-28
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there have been reports on the total protein two-dimensional electrophoresis of tomato and Arabidopsis, but the extraction and purification method of the total protein of the aquatic plant Zizania zizania by solid-phase two-dimensional electrophoresis has not been reported yet.

Method used

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  • Method for extracting and purifying wild rice stem total proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Interactive culture

[0033] (1) Isolation and culture of wild black powder fungus:

[0034] Pick a small amount of teliospores from the fresh cut surface of ash, and spread them evenly on a PSA plate (preparation: potato (peeled) 200g, sucrose 20g, agar 20g, add water to 1000mL, natural pH), culture 4d. Pick the characteristic colony and inoculate it on the PSA plate again after purifying and culturing for 18 days, punch holes on the edge of the active colony with a 4-5mm puncher, and inoculate the circular bacterial block on the PSA plate (10ml per dish, the diameter of the dish is 90mm, pH6 .0) on. Other culture conditions are: 24-28°C, relative humidity 75-85%.

[0035] (2) Tissue culture of wild rice:

[0036] The alternate lateral buds of the male water chestnut stem were used as the culture material, inoculated on the callus induction medium (preparation of the callus induction medium: MS medium 1000mL (Qingdao Hi-Tech Park Haibo Biotechnology Co.,...

Embodiment 2

[0039] Example 2: Extraction of Zizania total protein

[0040] (1) Weigh about 3 g of Zizania zizania callus after 12 days of interactive culture, cut it into pieces, add 0.3 g of tissue abrasive PVP-40 (Amresco) and a small amount of quartz sand, and grind it into a powder (gray green) with liquid nitrogen.

[0041] (2) Put the ground wild rice stem sample into a 50ml centrifuge tube with a stopper, add 10ml of acetone solution with a concentration of 10% trichloroacetic acid (w / w) and a concentration of β-mercaptoethanol of 0.07% (w / w), and shake slightly , placed at -20°C for 1 h.

[0042] (3) Place the stoppered centrifuge tube in an ultracentrifuge at 4° C. and 13,000 rpm, and centrifuge for 20 minutes (the supernatant is dark green, and the precipitate is light green).

[0043] (4) Pour off the supernatant, add 10ml of acetone solution containing β-mercaptoethanol concentration of 0.07% (w / w), shake, and place it at -20°C for precipitation for 1h. (The supernatant is li...

Embodiment 3

[0049] Example 3: Extraction of Zizania zizania total protein

[0050] (1) Weigh about 3 g of Zizania zizania callus that has been cultured for 15 days, cut it into pieces, add 0.3 g of tissue abrasive PVP-40 and a small amount of quartz sand, and grind it into a powder (gray green) with liquid nitrogen.

[0051] (2) Put the ground wild rice stem sample into a 50ml centrifuge tube with stopper, add 9.6ml of acetone solution with a concentration of 10% trichloroacetic acid and 0.07% concentration of β-mercaptoethanol, shake slightly, and place at -20°C for precipitation for 1.5h .

[0052] (3) Place the stoppered centrifuge tube in an ultracentrifuge at 4° C. and 13,000 rpm, and centrifuge for 20 minutes (the supernatant is dark green, and the precipitate is light green).

[0053] (4) Pour off the supernatant, add 9.8ml of 0.07% β-mercaptoethanol in acetone solution, shake, and place at -20°C for precipitation for 2 hours (the supernatant is light green or green, and the preci...

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Abstract

The invention provides an extraction method of zizania latifolia protein in the interaction of zizania latifolia and zizania smut, which simulates the interaction between zizania latifolia and zizania smut, to extract zizania latifolia gene groups and proteins which are not mixed. The invention can prepare the zizania total protein product without the protein of zizania smut, whose protein content is high, impurity content is low and nucleic acid content is low, thereby satisfying the demand of two-dimensional electrophoresis to protein purity.

Description

(1) Technical field [0001] The invention relates to a method for extracting and purifying the total protein of water-white plants by simulating the mutual cultivation of water-white and black powder fungi, in particular to a method for extracting the total protein of water-white plants without contamination with wild black powder bacteria, which can be used for the extraction of water-white plant total protein by solid-phase two-dimensional electrophoresis. purification method. (2) Background technology [0002] Water bamboo (zizania latifolia Turcz), also known as water bamboo shoots, is a perennial aquatic perennial herb, asexual reproduction, native to China and Southeast Asia, and is an important aquatic vegetable in my country. The variety resources of water bamboo shoots in my country are quite rich, and it is the second largest aquatic vegetable species after lotus root. Its fleshy stems contain protein, fat, carbohydrates, crude fiber, etc. Before water bamboo shoots...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14C07K1/30
Inventor 叶子弘邹克琴俞晓平刘倩尤文雨
Owner CHINA JILIANG UNIV
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