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Parietochloris incisa delta 6 FAD gene promoter and application thereof

A promoter and gene technology, applied in the field of genetic engineering, can solve the problem of low product efficiency

Active Publication Date: 2015-12-30
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the substrate preference analysis of the Δ6FAD of the notch edge green algae, it is found that Δ6FAD can not only convert LA into GLA, but also catalyze ALA into SDA, but the efficiency of the product is not high, which also illustrates the rate-limiting effect of Δ6FAD

Method used

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  • Parietochloris incisa delta 6 FAD gene promoter and application thereof
  • Parietochloris incisa delta 6 FAD gene promoter and application thereof
  • Parietochloris incisa delta 6 FAD gene promoter and application thereof

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Embodiment 1

[0038] The main operations adopted by the present invention include:

[0039] (1) Inoculate the Erlenmeyer flask of BG-11 medium with the green algae inoculated into the conical flask of BG-11 medium, and place it at a temperature of 25°C and a light intensity of 115 μmolphotonsm. -2 the s -1 1. Cultivate in an incubator with light cycle: light: dark / 12h:12h, shake regularly several times a day. Algae somatic cells were collected and total genomic DNA was extracted.

[0040] (2) Design gene-specific primers (GSP1: SEQ ID NO.1; GSP2: SEQ ID NO.2; GSP3: SEQ ID NO.3) according to the full-length cDNA sequence of the Δ6FAD gene of Chlorella nicked margin (Accession No. JN205755), and clone it by genome walking technology The 5'-flanking sequence (SEQ ID NO.4) upstream of the Δ6FAD gene was obtained, and then the regulatory elements of the promoter sequence were predicted online through a web server.

[0041] (3) According to the obtained Δ6FAD gene upstream 5'-flank sequence an...

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Abstract

The invention relates to a Parietochloris incisa delta 6 FAD gene promoter and an application thereof, and in particular provides a Parietochloris incisa delta 6 FAD gene promoter sequence, an application of the promoter sequence in promoting algae expression target genes, and a recombinant vector containing the promoter sequence and genetically engineered host cells. An obtained Parietochloris incisa delta 6 FAD gene promoter is connected into a fusion expression vector by cloning, a saccharomyces cerevisia BY4741 is transformed by an electric shock transformation method, the promoter is found to be capable of driving the expression of a target gene, and the effective rate of the promoter is almost equivalent to that of a T7 promoter. The Parietochloris incisa delta 6 FAD gene promoter provided by the invention creates a condition for artificial regulation and control of expression of genes of alga desaturase and the like.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a promoter of the Δ6FAD gene of a notch-margined green algae and its application. Background technique [0002] Arachidonic acid (ArA, 20:4ω6) is an essential fatty acid for human nutrition. Not only has many physiological and pharmacological activities, but also is the main component of human meningeal phospholipids. It plays the role of 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 alkenes. ArA is also an important component of mature breast milk and is necessary for the visual and intellectual development of postpartum infants. Therefore, FAO / WHO recommends adding ArA to infant formula. Myrmecia incisa is rich in a large amount of ArA, especially under stress conditions (such as phosphorus starvation, ni...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/80C12N1/15
Inventor 周志刚张利宁璞
Owner SHANGHAI OCEAN UNIV
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