Deoxyribose nucleic acid (DNA) sequence for encoding parietchloris incise diacylglycerol acyltransferase and application thereof

A technology of diacylglycerol acyl group and DNA sequence, which is applied in the field of genetic engineering, and can solve the problems of undisclosed cDNA sequence and errors of diacylglycerol acyltransferase from Chlorella diacylglycerol

Active Publication Date: 2013-02-27
SHANGHAI OCEAN UNIV
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Problems solved by technology

However, the paper did not disclose the cDNA sequence of the diacylglycerol acyltransferase of Chlorella nicked margin, and those skilled in the art know that there are errors in the high-throughput sequencing method, and the measured cDNA sequence is not 100% accurate, and there may be errors Great sex

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  • Deoxyribose nucleic acid (DNA) sequence for encoding parietchloris incise diacylglycerol acyltransferase and application thereof
  • Deoxyribose nucleic acid (DNA) sequence for encoding parietchloris incise diacylglycerol acyltransferase and application thereof
  • Deoxyribose nucleic acid (DNA) sequence for encoding parietchloris incise diacylglycerol acyltransferase and application thereof

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Embodiment 1

[0052] 1. Experimental materials

[0053] 1) Notched margin green algae ( Myrmecia incisa Reisigl H4301) were purchased from the Center for Algae Culture (CAUP), Charles University, Prague, Czech Republic. At a temperature of 25°C and a light intensity of 115 μmol photons m -2 ·s -1 , cultivated under the condition of light / dark ratio of 12h / 12h, medium BG-11, purchased from Shanghai Huyu Biotechnology Co., Ltd.

[0054] 2) pMD19T vector (vector map see figure 1 ), pYES2 vector (vector map see figure 2 ) were purchased from Invitrogen. Yeast-deficient strain H1246 and wild-type scy62 were purchased from the laboratory of Dr. Stymne, Swedish University of Agricultural Science and Technology. Yeast was cultured in SC medium at 30°C with shaking at 200 rpm. SC medium was purchased from Shanghai Enzyme Biotechnology Co., Ltd. Escherichia coli DH5α competent cells were purchased from Shanghai Sangon Biological Co., Ltd. The oil body-specific fluorescent dye BODIPY for l...

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Abstract

The invention relates to a separated deoxyribose nucleic acid (DNA) sequence. The DNA sequence comprises a nucleotide sequence (a) of SEQIDNO.1 or SEQIDNO.2; or a nucleotide sequence (b) complementary with the nucleotide sequence (a). The invention further provides a recombinant expression vector containing the nucleotide sequence, a genetic engineering host cell and application of the nucleotide sequence, the recombinant expression vector and the host cell for encoding the diacylglycerol acyltransferase or producing triacylglycerol. The DNA sequence has the advantages that a cDNA full-length sequence and a DNA full-length sequence of a parietchloris incise diacylglycerol acyltransferase gene are obtained by screening, encoded protein of the gene has a triacylglycerol synthesis capability by expression of the gene in a TAG synthesis defect strain H1246 of yeast, and conditions are created for utilizing the gene to achieve large-scale synthesis of triacylglycerol by genetic operation.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to a DNA sequence encoding Diacylglycerol Acyltransferase (MiDGAT2) from Chlorella nickedlis and its application. Background technique [0002] Energy is the basis for the development of human society. At present, the world is facing an energy crisis and the supply of oil resources is tight. Due to the renewable nature of bio-energy, the development of bio-energy has attracted more and more attention, and microalgae biodiesel has attracted more and more attention of many researchers. [0003] Different species of microalgae vary in their ability to synthesize triacylglycerol (TAG). Studies have found that eukaryotic microalgae such as green algae and diatoms have relatively high oil content, and they are expected to become biodiesel production algae species. Notch edge green algae ( Myrmecia incisa Reisigl H4301) is a freshwater unicellular microalgae belong...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12P7/64C12N15/63C12N1/21C12N1/15C12N1/19C12N5/10C12R1/89
Inventor 周志刚房逢立
Owner SHANGHAI OCEAN UNIV
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