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Lysophosphatidic acid acyltransferase gene and its application in notch-margined green algae

A technology of oil and nucleotides, applied in the field of bioenergy and plant biology

Active Publication Date: 2019-09-06
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, there have been no reports on the cloning of the lysophosphatidic acid acyltransferase gene from notch-margined green algae, especially the lysophosphatidic acid acyltransferase gene from notch-margined green algae with high utilization value.

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Embodiment Construction

[0035] The specific embodiments provided by the present invention will be described in detail below in conjunction with the accompanying drawings.

[0036] The main operation steps that the present invention adopts comprise:

[0037] 1) At a temperature of 25°C and a light intensity of 115μmol photons / (m 2 · s), under the condition that the light-to-dark ratio is 12h / 12h, cultivating green algae in the BG-11 medium. Algal cells were collected, and total RNA was extracted using the TRIzol method.

[0038] 2) Based on the transcriptome database of notch margin green algae (Ouyang et al., 2013) obtained by our laboratory, through the BLASTx comparison of NCBI, it was found that contig10781 and Coccomyxa subellipsoidea (Coccomyxa subellipsoidea) C-169 partial LPAAT sequence (GenBank accession No.: XP_005650277) has 70% similarity. Primers were designed according to this sequence, and the sequence of contig10781 was first cloned and verified using reverse transcription-PCR (RT-P...

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Abstract

The invention relates to a myrmecia incisa lysophosphatidic acid acyltransferase (LPAAT) gene and application thereof. According to the invention, the gene of myrmecia incisa LPAAT, which is considered to be a key enzyme for TAG synthesis, is cloned and is named as MiLPAAT; the prokaryotic expression vector of MiLPAAT is further constructed; after escherichia coli BL21 transformation and induced culture, the MiLPAAT is purified to obtain MiLPAAT mature protein with recombinant expression; the in vitro enzyme activity assay and the thin-layer chromatography result of enzyme-catalyzed reaction product show that the recombinant MiLPAAT mature protein can be used to synthesize PA from LPA, indicating that the protein encoded by MiLPAAT has the function of LPAAT to catalyze the synthesis of PAfrom LPA, and PA can be further used to synthesize TAG in algae cells along the Kennedy pathway. The application lays the foundation for overexpression of the MiLPAAT gene in grain and oil crops to increase the yield of triacylglycerol.

Description

technical field [0001] The invention relates to the fields of bioenergy and plant biotechnology, in particular to a gene of a notch-margined green algae lysophosphatidic acid acyltransferase (LPAAT) and its application. Background technique [0002] At present, various mineral resources such as coal, oil, natural gas, etc. are gradually exhausted with continuous development and utilization, and human beings urgently need to find new energy sources, especially renewable energy sources. Scientists believe that bioethanol and biodiesel are two important liquid biofuels, and it is possible to replace petroleum-based fuels through large-scale production of biomass. Biodiesel has become a hotspot in the development of new energy sources all over the world because of its advantages such as wide sources, degradability, high calorific value and environmental friendliness. Among them, the use of microalgae to produce biodiesel has become the focus of increasing attention of scientist...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/82A01H5/00
Inventor 周志刚常依娜包虹
Owner SHANGHAI OCEAN UNIV