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Genes with triacylglycerol synthesis function and their application in rational regulation of triacylglycerol content or saturation in oleaginous microalgae
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A technology of triacylglycerol and gene, applied in application, genetic engineering, acyltransferase, etc., can solve problems such as low oil yield and oil saturation that has not yet reached the application standard
Active Publication Date: 2021-08-27
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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Problems solved by technology
However, at present, microalgae oil production also faces many bottlenecks, such as low oil yield, and oil saturation has not yet reached the application standard, etc.
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Embodiment 1
[0045] Example 1: Cloning and analysis of NoDGAT gene
[0046] The NoDGAT2A, 2C and 2D genes and their flanking sequences were respectively cloned from the gDNA and cDNA of IMET1 by PCR technology. Restriction sites, synthesized by Shanghai Sangong:
[0047] NoDGAT2A primer pair is
[0048] 1) NoDGAT2A-for:
[0049] 5'GGTACCACATAATGACGCCGCAAGCCGAC3';
[0050] 2) NoDGAT2A-rev:
[0051] 5'GAATTCTTACTCAATGGACAACGGGCGCGTCT 3';
[0052] 3) NoDGAT2C primer pair is NoDGAT2C-for:
[0053] 5'GGTACCACATAATGACATCCTCCCCACC 3';
[0054] 4) NoDGAT2C-rev:
[0055] 5'GAATTCTCACCTGACCACTAAGGTGGCC 3';
[0056] NoDGAT2D primer pair is
[0057] 5) NoDGAT2D-for:
[0058] 5' GGTACCACATAATGAAGAAAATCTTGCGC 3';
[0059] 6) NoDGAT2D-rev:
[0060] 5'GAATTCCTAATAAAGCTCCAGCTCCCTGT 3'.
[0061] The PCR machine used was MasterCycler from Eppendorf Company, the reaction system was 50 μL, including 4 μL dNTP (2.5 mM each, TAKARA), 2 μL each of forward and reverse primers (10 μM), 5 μL 10×buffer (...
Embodiment 2
[0064] Example 2: Overexpression of NoDGAT in marine Nannochloropsis IMET1
[0065] (1) Construction of endogenous overexpression vector
[0066] see figure 2 . The specific method for constructing the vector is as follows: the full-length ORF fragment of NoDGAT is amplified by using the cDNA of Nannochloropsis IMET1 as a template by PCR method. In order to construct the cloning, with the help of primer introduction method, a restriction site and a protective base are added to the 5' end of the target sequence, and a restriction site is added to its 3' end. The sequence of the primer pair is as follows:
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Abstract
The invention belongs to the field of biotechnology. The invention relates to a gene with triacylglycerol (TAG) synthesis function and the application of the gene in increasing the TAG content and rationally regulating the TAG saturation of microalgae. The gene is the base sequence shown in SEQ ID NO 1, the base sequence shown in SEQ ID NO2 or has the base sequence shown in SEQ ID NO3; and, respectively with the base sequence shown in SEQ ID NO 1, SEQ ID The base sequence shown in NO 2 or the base sequence shown in SEQ ID NO 3 has more than 95% homology and encodes a DNA sequence of the same biological function protein. The genes of the present invention are three diacylglycerol acyltransferase DGAT genes isolated from Nannochloropsis. The content and saturation of TAG can be rationally regulated by using the obtained gene, in view of the fact that biodiesel, health products and medicines have different demands on the saturation of intracellular triacylglycerol.
Description
technical field [0001] The invention belongs to the field of biotechnology. The invention relates to a gene with triacylglycerol (TAG) synthesis function and the application of the gene in increasing the TAG content and rationally regulating the TAG saturation of microalgae. Background technique [0002] Under the guidance of the concepts of energy saving, emission reduction, and low carbon, the development of renewable energy has become an important research direction. At present, most renewable energy sources provide energy in the form of power generation, but since most transportation vehicles still use liquid fuels as energy sources, electric energy cannot directly solve the power problem of current transportation vehicles. Biofuel is currently the only renewable energy that can be used as liquid fuel, so it occupies an important position in the development of renewable energy. Among them, biodiesel is most suitable for the current internal combustion engine system, wh...
Claims
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