Polypeptide for improving triacylglycerol synthesizing capability of MiDGAT1 in beer yeast and application of polypeptide

A technology of yeast cells and host cells, applied in the field of genetic engineering

Active Publication Date: 2015-12-09
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has been proved that DGAT1 has the function of TAG synthesis in many higher plants, there are still relatively few studies on the function of TAG synthesis by microalgae DGAT1. The functional study of the PH domain has not been reported yet

Method used

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  • Polypeptide for improving triacylglycerol synthesizing capability of MiDGAT1 in beer yeast and application of polypeptide
  • Polypeptide for improving triacylglycerol synthesizing capability of MiDGAT1 in beer yeast and application of polypeptide
  • Polypeptide for improving triacylglycerol synthesizing capability of MiDGAT1 in beer yeast and application of polypeptide

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Experimental program
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Embodiment 1

[0045] 1. Materials

[0046] (1) Notched margin green algae (Myrmecia incisa Reisigl H4301) were purchased from the Algae Culture Center (CAUP) of Charles University in Prague, Czech Republic. The algae were inoculated in a conical flask containing BG-11 medium, placed at a temperature of 25 °C and a light intensity of 115 μmolphotons m -2 ·s -1 1. Cultivate in an incubator with light cycle: light: dark / 12h:12h, shake regularly several times a day.

[0047] (2) The pYES2 vector was purchased from Invitrogen. Wild-type brewer's yeast (Saccharomycescerevisiae) Scy62 and yeast-deficient strain H1246 were purchased from the laboratory of Dr. Stymne, Swedish University of Agricultural Science and Technology. Yeast was inoculated in SC medium, and cultured with shaking at 200 revolutions per minute (rpm) at 28°C.

[0048] 2. Method

[0049] (1) Take 100 mg of fresh algae cells and put them into a pre-cooled mortar, add liquid nitrogen and grind them thoroughly.

[0050] (2) Us...

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Abstract

The invention relates to polypeptide for improving the triacylglycerol synthesizing capability of MiDGAT1 in beer yeast and application of the polypeptide. Particularly, a new DGAT gene is obtained through cloning and named MiDGAT1, a new PH structural domain sequence is obtained, it is found through a gene functional complementation experiment that proteins coded by MiDGAT1 and MiDGAT1-deltaPH have the TAG synthesizing capability, however, it is found through a semi-quantitative thin-layer chromatography method that the content of TAG synthesized through MiDGAT1 yeast is much higher than that synthesized through MiDGAT1-deltaPH yeast, and therefore it is proved that the PH structural domain sequence can improve the capability of the MiDGAT1 in synthesizing the TAG in the yeast, and the improving function of the PH structural domain sequence is obviously higher than that of a PH structural domain sequence in the prior art. Accordingly, MiDGAT1 containing a PH structural domain can be expressed in crops such as oilseeds to obviously increase the yield of triacylglycerol.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the identification of the Pleckstrin homology (Pleckstrin Homology, PH) domain (domain) sequence and function of Chlorella diacylglycerol acyltransferase 1 (MiDGAT1). Background technique [0002] In today's world, with the continuous depletion of various mineral resources, such as coal, oil, and natural gas, human beings are eager to find new energy sources, especially renewable resources. Biodiesel, especially microalgae biodiesel, is a renewable resource, one of the new energy sources that human beings are eager to find and explore, and it is also one of the research hotspots of scientists in recent years. Compared with traditional biodiesel raw materials, such as vegetable oil and animal fat, microalgae have many advantages such as fast growth, rich biomass, high oil content and no land occupation. Notched margin green algae (Myrmecia incisa Reisigl H4301) is a f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12P7/64
CPCC12N9/1029C12P7/6463C12Y203/0102
Inventor 周志刚陈春秀
Owner SHANGHAI OCEAN UNIV
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