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A method for rapid expression of foreign proteins in plants

An exogenous protein and plant technology, which is applied in botany equipment and methods, biochemical equipment and methods, plant products, etc., can solve the problems of inapplicable scale production of a large number of plant exogenous proteins, and achieve easy promotion and application, low cost Low cost, simple and quick operation

Inactive Publication Date: 2018-06-19
JILIN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main infection methods are leaf injection method and vacuum infection method, but neither of these two methods is suitable for the infection of a large number of plants and the large-scale production of exogenous proteins.

Method used

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  • A method for rapid expression of foreign proteins in plants
  • A method for rapid expression of foreign proteins in plants
  • A method for rapid expression of foreign proteins in plants

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0031] Using different concentrations of bacterial solution, MMA OD 600 Infection was carried out at 0.6, 0.8, 1.0, 1.2, and 1.4, respectively. The seedlings at the four-leaf stage of Nb plants (about 18 days after transplanting) were used as host materials, MMA OD 600 The efficiency of GFP expression is 65% when it is 0.6; MMA OD 600 The efficiency of GFP expression is 75% when it is 0.8; MMA OD 600 The efficiency of GFP expression is 90% at 1.0; MMA OD 600 The efficiency of GFP expression was 82% at 1.2; MMA OD 600 The efficiency of GFP expression was 60% at 1.4; MMA OD 600 When it is 1.6, the efficiency of GFP expression is 45%. The expression efficiency is determined by the percentage of the plants with the green fluorescence phenomenon phenotype in the total number of plants (20 plants at each concentration). The experiment is repeated three times, and the percentages are calculated respectively and averaged (such as figure 2 ). The results of the study showed tha...

example 2

[0033]Sow the seeds of Nb in 1 / 2MS medium (searched on the Internet), put them in a light incubator, and cultivate them for about a week. The seedlings were transplanted into flowerpots, and cultivated under the condition of temperature of 25±3°C, under the cycle condition of 16 hours of light and 8 hours of darkness every day, and the seedlings growing for 18-28 days were used as infection materials. With the seedlings at the four-leaf stage of Nb plants as the host material, the efficiency of GFP expression was 90%; with the seedlings at the five-leaf stage as the host material, the efficiency of GFP expression reached 75%; with the seedlings at the six-leaf stage as the host material, GFP The efficiency of expression was 55%. The expression efficiency is determined by the percentage of the plants with the green fluorescence phenomenon phenotype in the total number of plants. The experiment is repeated three times, and the percentages are respectively calculated and averaged...

example 3

[0035] The expression efficiency of GFP was also influenced by the incubation temperature of the plant material after infection. Put the infected tobacco seedlings in a light incubator and cultivate them for about a week. The culture is carried out under the cycle conditions of 16 hours of light and 8 hours of darkness every day. At 22°C, the efficiency of GFP expression is 65%; at 25°C, the efficiency of GFP expression is 80%; at 28°C Under the condition, the efficiency of GFP expression is 92%; under the temperature condition of 31°C, the efficiency of GFP expression is 75%; under the temperature condition of 34°C, the efficiency of GFP expression is 60%. The expression efficiency is represented by the percentage of the plants with the green fluorescence phenotype in the total number of plants. The experiment was repeated three times, and the percentages were calculated and averaged (such as Figure 4 ). The results showed that the culture temperature of plant materials af...

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Abstract

The invention belongs to a method for rapidly expressing foreign protein in plants. The method comprises the following steps: 1. preparation of plant material, 2. construction of virus vector, 3. preparation of agrobacterium suspension, 4. infection of plant using high-pressure spray gun system, 5. cultivation of plant material after infection and Observation, ⑥, rapid expression and detection of GFP. The invention is a new method for expressing exogenous protein conveniently and rapidly in plants, and the green fluorescent protein GFP is successfully expressed by using the method. In a large number of optimization experiments, the applicant conducted a relatively comprehensive and specific research on various factors affecting gene expression, and determined the optimal infection conditions of the method, including the concentration of the bacterial liquid resuspension (MMA OD600), and the plants suitable for the method Material and infection period, temperature conditions for plant material cultivation, etc.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for rapidly expressing foreign protein in plants. Background technique [0002] In recent decades, a large number of studies have shown that plant bioreactors can effectively express many foreign proteins, including human serum proteins, growth factors, antibodies, vaccines, etc. At present, medicinal proteins produced by plants abroad have been used in clinical practice, and plant bioreactors are increasingly showing their importance. There are mainly two expression systems in plant bioreactors, namely stable expression system and transient expression system. The plant transient expression system is more and more favored by people because of its fast, safe and effective advantages, and has gradually become one of the domestic research hotspots. Its practical value and development prospects have been generally affirmed. [0003] The plant transient expression system is med...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/82
Inventor 杨丽萍金太成王艳
Owner JILIN NORMAL UNIV
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