Applications of grease metabolism related protein GmNF307 in plant grease metabolism regulation
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A plant and oil technology, applied in applications, plant peptides, plant products, etc., can solve problems such as little known
Active Publication Date: 2016-10-19
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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However, in plants, the regulatory mechanism of lipid synthesis and its associated genes are still poorly understood
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Embodiment 1
[0090] Example 1. Cultivation of Transgenic Arabidopsis Plants Using Lipid Metabolism-Related Protein Genes
[0091] This embodiment provides a soybean-derived lipid metabolism-related protein gene, which is named as the lipid metabolism-related protein GmNF307 gene.
[0092] Prepare the DNA molecule shown in SEQ ID No.1 in the sequence table (i.e. the lipid metabolism-related protein GmNF307 gene, referred to as GmNF307 gene), the DNA molecule shown in SEQ ID No.1 encodes the protein shown in SEQ ID No.2 (i.e. Lipid metabolism-related protein GmNF307, referred to as GmNF307 protein).
[0093] 1. Expression analysis of GmNF307 gene in different organs of soybean
[0094] 1.1. Extract the total RNA of the soybean globular embryo, reverse transcribe and synthesize cDNA with reverse transcriptase to obtain the soybean globular embryo cDNA.
[0095] 1.2, according to the method of 1.1, replace the soybean globular embryo in 1.1 with soybean heart-shaped embryo, soybean cotyledon...
Embodiment 2
[0116] Example 2, Phenotype analysis of transgenic Arabidopsis thaliana
[0117] 1. Determination of total oil content in seeds
[0118] The experiment was repeated three times, and the specific steps for each repetition were as follows:
[0119] Grind the dried seeds into powder, weigh 100 mg into a centrifuge tube, and weigh four portions in parallel. Add 500 μl of n-hexane, mix thoroughly, and leave overnight at 37°C. Slowly centrifuge for 3 minutes, and draw n-hexane into a new weighed tube. Continue to add n-hexane to the remaining powder to soak repeatedly, then centrifuge, and then collect n-hexane into the same centrifuge tube. Put the centrifuge tube into the vacuum pump and pump the vacuum to make the n-hexane completely volatilize. Then weigh the centrifuge tube again. The change in weight before and after the centrifuge tube is the total oil weight extracted; the formula for calculating the total oil amount (%) is:
[0120] Total oil content (%) = (weight of ...
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Abstract
The present invention discloses applications of a grease metabolism related protein GmNF307 in plant grease metabolism regulation. The protein is the following protein a) or b) or c): a) the protein has the amino acid sequence represented by SEQ ID No.2; b) the fusion protein is obtained by linking a tag to the N terminal or / and C terminal of the protection represented by SEQ ID No.2; and c) the protein is obtained by substituting and / or deleting and / or adding one or a plurality of amino acid residues on the amino acid sequence represented by SEQ ID No.2 and is associated with the grease metabolism regulation. According to the present invention, the experiment results show that the contents of the total greases in plant tissues and / or organs and the contents of partial fatty acids in plant tissues and / or organs can be increased with the overexpression of the encoding gene GmNF307 of the grease metabolism related protein GmNF307.
Description
technical field [0001] The invention relates to the application of lipid metabolism-related protein GmNF307 in the regulation and control of vegetable lipid metabolism in the field of biotechnology. Background technique [0002] Among several major oil-producing crops in the world, soybean oil production accounts for about 30%, ranking first in the world's vegetable oil production. [0003] The synthesis of fatty acid is one of the most important metabolic pathways in plants. It exists in any cell of the plant and is necessary for growth and development. Blocking its synthesis will lead to cell death, so it has not been found so far. A plant mutant that blocks fatty acid synthesis. The enzymes involved in the fatty acid synthesis pathway differ greatly between plants and other eukaryotes. Synthesis of fatty acids with 16 or 18 carbon atoms from acetyl CoA and malonyl CoA requires at least 30 different enzyme-catalyzed reactions to complete this process, and in animals, fun...
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