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A method for the identification of fatty acid desaturase genes associated with DHA biosynthesis in Cryptodinium algae using de novo transcriptome analysis

A technology of biosynthesis and desaturase, applied in biochemical equipment and methods, microbial measurement/testing, genomics, etc., can solve the problems of unclear synthesis pathway of DHA and restriction of Cryptidium korii

Active Publication Date: 2020-10-30
KUNMING ZAONENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current genome information of Cryptidium is not published, and its DHA synthesis pathway is still unclear, which limits various efforts to engineer C.

Method used

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  • A method for the identification of fatty acid desaturase genes associated with DHA biosynthesis in Cryptodinium algae using de novo transcriptome analysis
  • A method for the identification of fatty acid desaturase genes associated with DHA biosynthesis in Cryptodinium algae using de novo transcriptome analysis
  • A method for the identification of fatty acid desaturase genes associated with DHA biosynthesis in Cryptodinium algae using de novo transcriptome analysis

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Embodiment 1

[0058] A method for identifying a fatty acid desaturase gene related to DHA biosynthesis in Cryptodinium, comprising the steps of:

[0059] (1) Carry out the fermentative culture of DHA synthesis to Cryptidinium:

[0060] Cryptidium was cultured and grown statically at 25° C. for 3-5 days in a medium containing 2.0 g / L yeast extract, 9.0 g / L glucose, and 25.0 g / L seawater salt. Then with 10% (v / v) bacterial classification inoculum, inoculate in the 250 milliliters conical flasks that contain 50mL medium (2.0g / L yeast extract, 25.0g / L sea salt and 9.0g / L glucose), And cultivated on a shaker for 2 days (180 rpm, 25° C.). Subsequent scale-up inoculation into 500-mL Erlenmeyer flasks containing 100 mL of medium (7.5 g / L yeast extract, 25.0 g / L sea salt, 27.0 g / L glucose) was used to inoculate 7.5 L of Bioreactor. Finally transfer to fermentor culture (7.5g / L yeast extract, 25.0g / L sea salt, 27.0g / L glucose) with 20% (v / v) strain inoculum amount, and according to C:N in the firs...

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Abstract

The invention discloses a method of identifying DHA (docosahexaenoic acid) biosynthesis related fatty acid desaturase gene in crypthecodinium cohnii through De novo transcriptome sequencing, transcript splicing and differential expression analysis. The method particularly includes the following steps: (1) performing DHA fermentation culture to the crypthecodinium cohnii; (2) collecting thallus at different growth and DHA accumulation stages, and performing RNA extraction and transcriptome Illumina deep sequencing; (3) analyzing and pre-processing sequenced data; (4) performing function annotation and correlation statistic analysis to the spliced transcriptome, identifying the DHA biosynthesis related fatty acid desaturase in the crypthecodinium cohnii and acquiring a whole gene sequence thereof; and (5) by means of qRT-PCR analysis technology, performing expression verification to the DHA biosynthesis related fatty acid desaturase gene in the crypthecodinium cohnii. The method provides important approaches and knowledge basis for researching biosynthesis mechanism of DHA in the crypthecodinium cohnii and improving DHA synthesis capability of the crypthecodinium cohnii.

Description

technical field [0001] The invention belongs to the field of biological food, and relates to a method for identifying a fatty acid desaturase gene related to DHA biosynthesis in Cryptodinoflagellates and an application thereof. Background technique [0002] DHA (Docosahexaenoic Acid, DHA) is one of the important substances for human brain development and growth, and is an important component of the brain and retina. The largest proportion, accounting for about 50%. Therefore, it plays an important role in the development of the brain and retina of fetuses and infants, as well as the maintenance of adult brain functions. In addition, DHA also has a positive effect on diseases of the cardiovascular system such as coronary heart disease, stroke, hypertension, nervous system diseases such as neurological disorders, Alzheimer's disease and depression. As an essential fatty acid, DHA has more significant effects on enhancing memory and thinking ability and improving intelligence...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C40B50/06G16B20/00G16B20/30G16B25/20
CPCC12Q1/6806C12Q1/6869C40B50/06G16B20/00G16B30/00C12Q2535/122
Inventor 张卫文陈磊裴广胜刘璐
Owner KUNMING ZAONENG BIOTECH CO LTD