DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase and application thereof

A technology of fatty acid elongation enzyme and notch edge green algae, which is applied in the field of DNA sequence to achieve the effect of improving quality

Active Publication Date: 2012-11-21
SHANGHAI OCEAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the DNA sequence encoding Δ-6 fatty acid elongase and its application in green algae.

Method used

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  • DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase and application thereof
  • DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase and application thereof
  • DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase and application thereof

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Experimental program
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Embodiment

[0030] 1. Materials

[0031] 1) Notched margin green algae ( Myrmecia incisa Reisigl H4301) from CAUP (Culture collection of algae of Charles University of Prague). At a temperature of 25°C and a light intensity of 115 μmol photons m -2 the s -1 , The light / dark ratio is 12h / 12h and the culture medium is BG-11.

[0032] 2) pYES2 vector and yeast INVSc1 strain (His - 、Leu - 、Trp - and Ura - Defective type) was purchased from Invitrogen. Yeast was cultured in YPD medium (yeast extract, peptone, glucose medium) at 30°C with shaking at 220 rpm.

[0033] 2. Method

[0034] 1) Take 100 mg of fresh algae cells and place them in a pre-cooled mortar, add liquid nitrogen and grind them thoroughly.

[0035] 2) Genomic DNA was extracted by CTAB method (cetyltrimethylammonium bromide method) and total RNA was extracted by TRIzol method, and stored at -20°C for later use.

[0036] 3) Design primers according to the expressed sequence tag (EST) of the elongase gene screened in t...

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Abstract

The invention relates to a DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase, which comprises: a, the nucleotide sequence in SEQIDNO.10; or b, the nucleotide sequence complementary to the nucleotide sequence in a. The invention also provides a recombinant expression vector and genetically engineered host cells of the nucleotide sequence and the application thereof. The advantages of the invention are as follows: it is testified through the molecular cloning and functional identification of the Myrmecia incisa delta-6 fatty acid elongase gene that the gene code is delta-6 fatty acid elongase; the application provided by the invention provides a theoretical basis for improving the capability of long-chain polyunsaturated fatty acid synthesis from oil crops, and theoretically, the gene provided by the invention can be introduced into oil crops to effectively prolong the precursors that are necessary for the synthesis of arachidonic acid such as gamma-linolenic acid and endow the oil crops with the capability of the long-chain polyunsaturated fatty acid synthesis so as to improve the quality of the oil crops.

Description

technical field [0001] The present invention relates to a DNA sequence, specifically, a DNA sequence encoding the Δ-6 fatty acid elongase of green algae of the nick margin and its application. Background technique [0002] ArA (arachidonic acid, arachidonic acid, 20:4n-6) is an essential fatty acid for human nutrition, and it is the same as docosahexaenoic acid (DHA, 22:6n-3). The main component of meningeal phospholipids, as well as many other physiological and pharmacological activities. For example, it acts as a second messenger in cells, participates in the regulation of hematopoietic immunity, causes vasodilation, and participates in the regulation of various functions of liver, gallbladder and other organs. Precursors of trienes. ArA is also an important component of mature breast milk, which is necessary for the visual and intellectual development of postpartum babies. Therefore, FAO / WHO (1994) recommends adding ArA to infant formula. [0003] At present, the comm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/52C12N15/63C12N1/21C12N1/15C12N1/19C12N5/10C12P7/64C12R1/89
Inventor 周志刚于水燕刘士成李慧吴洪
Owner SHANGHAI OCEAN UNIV
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