A kind of recombinant plasmid including astaxanthin synthase fusion gene, recombinant bacteria and application

A technology of astaxanthin synthase and recombinant plasmid, which is applied in application, genetic engineering, recombinant DNA technology, etc., to achieve the effect of low cost, simple and fast method, and high efficiency

Active Publication Date: 2021-06-29
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no record and report on the successful expression of astaxanthin by using camelina in the prior art

Method used

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  • A kind of recombinant plasmid including astaxanthin synthase fusion gene, recombinant bacteria and application
  • A kind of recombinant plasmid including astaxanthin synthase fusion gene, recombinant bacteria and application
  • A kind of recombinant plasmid including astaxanthin synthase fusion gene, recombinant bacteria and application

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Embodiment 1

[0039] 1. Materials and methods

[0040] 1. Materials

[0041] Camelina sativa (L.) germplasm material SC-N16, (see [Yuan Lixia, Hao Jingyun, Zhao Kui, et al. 2EMS mutagenesis of Camelina sativa (L.) to obtain 2S storage protein deletion mutants. Journal of North China Agricultural Science, 2016, 31(3):11-17]), provided by the Key Laboratory of Crop Genetics and Breeding, Shanxi Agricultural University. Sow camelina seeds in seedling pots with a diameter of 9 cm, 1 seed per pot; the substrate in the pot is humus: peat soil: perlite = 5:4:1 (volume ratio); the temperature in the greenhouse is 16-22°C, and the light source is Natural light, 60% relative humidity. When the plants grow to 10cm, topping is carried out to promote the growth and development of side branches. When the plant grows for about 70 days, it enters the flowering stage and prepares for transformation, and the plant is thoroughly watered the day before the transformation. A total of 20 plants (pots) were t...

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Abstract

The invention provides a recombinant plasmid comprising an astaxanthin synthase fusion gene, a recombinant bacterium and its application, and belongs to the technical field of genetic engineering breeding. The invention connects four astaxanthin synthesis key enzyme genes with 2A short peptide sequence, and uses CaMV35S promoter to control expression, which can simplify the construction of recombinant plasmid (expression vector) and realize the coordinated expression of each gene. The recombinant plasmid of the present invention is introduced into camelina plants, and the seeds of the obtained camelina transformed plants synthesize and accumulate high-content carotenoids, wherein the content of astaxanthin increases from 0 to 15.23 μg / gDW. At the same time, the total antioxidant capacity of the seeds of the transformed strain was significantly higher than that of the wild type. Therefore, using the recombinant plasmid of the invention to breed camelina and producing high-quality edible oil with unsaturated fatty acids and carotenoids has great commercial prospects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and breeding, and in particular relates to a recombinant plasmid including an astaxanthin synthase fusion gene, a recombinant bacterium and its application. Background technique [0002] Astaxanthin is a red ester-soluble ketone carotenoid with a unique molecular structure, which has strong antioxidant activity, anti-radiation, anti-aging, anti-tumor and cardiovascular and other diseases prevention functions. The biosynthesis of astaxanthin only occurs in a small amount of bacteria and green algae, and the yield is very low; among plants, only Adonis aestivalis can synthesize trace amounts of astaxanthin in its petals. Due to the limited natural resources of astaxanthin, scientists are looking for different plants to produce higher yields of natural astaxanthin through genetic engineering methods, and in plants such as lettuce, potato, wheat, rapeseed, tobacco, tomato and rice. be suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N15/82C12N1/21A01H5/10A01H6/20C12R1/01
CPCC07K14/415C12N9/0069C12N9/0071C12N9/1085C12N15/825C12N15/8261C12Y205/01
Inventor 何明霞黄俊潮
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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