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Analysis method of microalgae proteome

An analysis method and proteome technology, applied in the field of biofuels, can solve problems such as seasons and environmental impacts

Active Publication Date: 2012-10-03
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of biodiesel is mainly through oil-producing plants, including various beans, etc. However, the growth of plants and the limitation of living environment make the oil production process seriously affected by seasons and the environment

Method used

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  • Analysis method of microalgae proteome
  • Analysis method of microalgae proteome
  • Analysis method of microalgae proteome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A method for analyzing the proteome of microalgae, comprising the steps of:

[0045] (1) Chlorella sorokiniana cell culture and collection:

[0046] Four different cell inoculation densities (1 × 10 4 cells / mL, 1×10 5 cells / mL, 1×10 6 cells / mL, 1×10 7 cells / mL (indicated by IN104, IN105, IN106, IN107 in turn) for culture, when the culture reaches OD560 of 0.8, take out the cultured microalgae liquid sample, centrifuge at 3000rpm, 4℃ for 3min, and collect the cells in the lower layer , after washing the cells with water, centrifuge at 3000rpm at 4°C for 3min, collect the cells in the lower layer and freeze-dry them at -80°C for 4h to obtain freeze-dried cells;

[0047] (2) Extraction of intracellular proteins:

[0048] ① Take 1 portion of each of the above four freeze-dried cells, 50 mg each, and place them in centrifuge tubes, add 0.5 mL of cell lysate to each tube, mix well, and intermittently sonicate on ice in the method of ultrasonication for 8 seconds and inte...

Embodiment 2

[0083] A method for analyzing the proteome of microalgae, comprising the steps of:

[0084] (1) Chlorella sorokiniana cell culture and collection:

[0085] Four different cell inoculation densities (1 × 10 4 cells / mL, 1×10 5 cells / mL, 1×10 6 cells / mL, 1×10 7 cells / mL (indicated by IN104, IN105, IN106, IN107 in turn) culture, when the culture reaches OD560 of 1.0, take out the cultured microalgae liquid sample, centrifuge at 4000rpm for 3min at 4°C, collect the cells in the lower layer, After washing the cells with water, centrifuge at 4000 rpm for 3 minutes at 4°C, and freeze-dry at -80°C for 5 hours to obtain freeze-dried cells;

[0086] (2) Extraction of intracellular proteins:

[0087] ① Take 1 portion of each of the above four types of freeze-dried cells, 80 mg each, and place them in centrifuge tubes, add 0.8 mL of cell lysate to each tube, mix well, and ultrasonicate intermittently on ice for 9 seconds and 5 seconds each time. A total of 50 seconds; add 10 μL of DN...

Embodiment 3

[0105] A method for analyzing the proteome of microalgae, comprising the steps of:

[0106] (1) Chlorella sorokiniana cell culture and collection:

[0107] Four different cell inoculation densities (1 × 10 4 cells / mL, 1×10 5 cells / mL, 1×10 6 cells / mL, 1×10 7 cells / mL (indicated by IN104, IN105, IN106, and IN107 in sequence) for culture, when the culture reaches OD560 of 1.5, take out the cultured microalgae liquid sample, centrifuge at 5000rpm, 4°C for 4min, and collect the cells in the lower layer. After washing the cells with water, centrifuge at 5000 rpm and 4°C for 4 minutes, collect the cells in the lower layer and freeze-dry them at -80°C for 6 hours to obtain freeze-dried cells;

[0108] (2) Extraction of intracellular proteins:

[0109] ① Take 1 portion of each of the above four types of freeze-dried cells, 100 mg each, and place them in centrifuge tubes respectively. Add 1 mL of cell lysate to each tube, mix well, and ultrasonicate intermittently on ice for 10 s ...

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Abstract

The invention discloses an analysis method of microalgae proteome. The analysis method comprises the steps of collecting microalgae cells; extracting intracellular protein; and identifying the differentially expressed proteins. The inventtion provides an approach for analyzing microalgae proteome. The approach involves cell collection, protein extraction and protein analysis, thereby obtaining protein qualitative and quantitative results of microalgae under different cultivation conditions, so as to provide the analysis method of microalgae proteome. The method has significance for promoting microalgae proteome research.

Description

field of invention [0001] The invention belongs to the field of biofuels, and relates to an analysis method of microalgae proteome. Background technique [0002] With the continuous development of human society, the demand for energy is increasing day by day, and the development of sustainable energy is particularly important in the case of increasingly depleted fossil fuel resources. The field of bioenergy is an emerging field of energy research, including bioethanol and biodiesel. Among them, the application of bioethanol has problems such as poor technical compatibility, which makes biodiesel have greater advantages. At present, the production of biodiesel is mainly through oil-producing plants, including various beans, etc. However, the growth of plants and the limitation of living environment make the oil production process seriously affected by seasons and the environment. Now, the production of diesel through microalgae has become a popular production route. Becaus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06G01N21/31
Inventor 元英进马倩陆姝欢吕亚金
Owner TIANJIN UNIV
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