Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation method of wheat complete chloroplasts

A chloroplast and wheat technology, applied in the field of preparation of complete wheat chloroplasts, can solve the problems of long time consumption, loss of chloroplast DNA, inability to obtain sufficient matrix protein, etc., and achieve the effect of short preparation time and simple procedure

Inactive Publication Date: 2016-08-31
HENAN AGRICULTURAL UNIVERSITY
View PDF6 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional separation and purification of chloroplasts often use sucrose density gradient centrifugation and differential centrifugation, which not only takes a long time, but the density gradient centrifugation alone takes more than 5 hours, and this method cannot distinguish between intact and damaged chloroplasts ( figure 1 B), inability to obtain sufficient matrix protein may lead to loss of chloroplast DNA, seriously affecting follow-up research

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method of wheat complete chloroplasts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Prepare Percoll gradient:

[0021] (1) Weigh 10 mg of erythorbic acid and glutathione (stored at 4°C) each and put into a 50 mL round-bottomed translucent polyethylene centrifuge tube;

[0022] (2) Add 15mL 2×GB (grinding buffer pH7.3, containing 0.1M K-Hepes, 0.66M sorbitol, 2mM MgCl 2 , 2mM MnCl 2 , 4mM Na 2 EDTA and 0.2% BSA, stored at 4°C) for dissolution;

[0023] (3) Add 15 mL Percoll (stored at 4°C) and mix well;

[0024] (4) Centrifuge at 4°C and 37,000g for 30 minutes (note: brake on). After the centrifugation is completed, carefully remove the centrifuge tube and place it on ice for later use.

[0025] 2. Chloroplast preparation (can be performed synchronously during centrifugation in step 1, and all operations are performed at low temperature):

[0026] (1) Vessel preparation: Take two 250mL Erlenmeyer flasks labeled GB and IB respectively, four 50mL round-bottomed centrifuge tubes, one beaker, one plastic funnel, and one pair of scissors and place th...

Embodiment 2

[0039] 1. Prepare Percoll gradient:

[0040] (1) Weigh 10 mg of erythorbic acid and glutathione (stored at 4°C) each and put into a 50 mL round-bottomed translucent polyethylene centrifuge tube;

[0041] (2) Add 15mL 2×GB (grinding buffer pH7.3, containing 0.1M K-Hepes, 0.66M sorbitol, 2mM MgCl 2 , 2mM MnCl 2 , 4mM Na 2 EDTA and 0.2% BSA, stored at 4°C) for dissolution;

[0042] (3) Add 15 mL Percoll (stored at 4°C) and mix well;

[0043] (4) Centrifuge at 4°C and 37,000g for 30 minutes (note: brake on). After the centrifugation is completed, carefully remove the centrifuge tube and place it on ice for later use.

[0044] 2. Chloroplast preparation (can be performed synchronously during centrifugation in step 1, and all operations are performed at low temperature):

[0045] (1) Vessel preparation: Take two 250mL Erlenmeyer flasks labeled GB and IB respectively, four 50mL round-bottomed centrifuge tubes, one beaker, one plastic funnel, and one pair of scissors and place them...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of biotechnology, and relates to the technical field of plant complete cell organ separation, in particular to a simple and rapid preparation method of wheat complete chloroplasts. The method comprises the following steps of 1, preparing a Percoll gradient; 2, preprocessing a wheat sample; 3, performing density gradient centrifuging on the wheat sample; 4, completing extraction of the chloroplasts. The Percoll is adopted as a medium for gradient centrifuging, wheat complete and damaged chloroplasts can be distinguished, the preparation time is short, the procedure is simple and convenient, the sufficient and complete chloroplasts are obtained, quantitative determination is performed, and then the chloroplasts can be precisely applied to extraction of chloroplast function protein, membrane protein, matrix protein and chloroplast DNA and various follow-up researches.

Description

technical field [0001] The invention belongs to the field of biological technology, relates to the technical field of separation and purification of complete plant organelles, and in particular relates to a simple and quick preparation method of complete wheat chloroplasts. Background technique [0002] Chloroplast is the most important organelle of plant cells. It is the place where plants carry out photosynthesis and nitrogen assimilation. It is the original source of all organic matter in nature. It is closely related to plant growth and crop yield. Obtaining complete chloroplasts is very important for studying chloroplast functions and chloroplasts. A necessary condition for soluble protein and membrane protein, chloroplast gene, etc. The traditional separation and purification of chloroplasts often use sucrose density gradient centrifugation and differential centrifugation, which not only takes a long time, but the density gradient centrifugation alone takes more than 5...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/00
Inventor 王小纯张浩然韦一昊熊淑萍马新明
Owner HENAN AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com