A combined expression vector and its application in the expression of astaxanthin in corn kernels

An expression vector, astaxanthin technology, applied in the biological field, can solve the problems of astaxanthin activity loss, fishy smell, and low content of active trans-astaxanthin, so as to increase the supply of precursor substances and high-efficiency biosynthesis Effect

Active Publication Date: 2020-12-29
捷康生物科技(海南)有限公司
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AI Technical Summary

Problems solved by technology

Although the content of astaxanthin produced by the above-mentioned microorganisms is much higher than that in the seeds of transgenic plants, the content of active trans-astaxanthin contained in them is low, and the astaxanthin is only in the cells, and it must be extracted before it can be used In industrial production, a large amount of astaxanthin activity is lost during the extraction process, and the yield is not necessarily higher than that of transgenic plant seeds, while transgenic corn seeds can be directly used for feed that requires astaxanthin without extraction, such as high-end aquaculture Salmon compound feed, rainbow trout feed, etc., can also be used for flamingo, poultry and pet feed, etc.
In addition, the astaxanthin derived from genetically modified corn has no peculiar smell, while the astaxanthin derived from algae has a fishy smell, and the astaxanthin derived from yeast has a fungal smell

Method used

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  • A combined expression vector and its application in the expression of astaxanthin in corn kernels
  • A combined expression vector and its application in the expression of astaxanthin in corn kernels
  • A combined expression vector and its application in the expression of astaxanthin in corn kernels

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Embodiment 1

[0026] Embodiment 1, maize haploid immature embryo identification method

[0027] 1. Construction of expression vector

[0028] After obtaining the sequence of the relevant functional gene in the public database NCBI, after optimizing the codon of the gene sequence, synonymously mutating the unnecessary enzyme cutting site, and then integrating it into the genetic transformation mediated by Agrobacterium by enzyme cutting ligation The intermediate vector pBDEN-mtp ( figure 1 ), forming a series of expression vectors pBDEN-CP-BZ, pBDEN-CP-BZ-LcyEi and pBDEN-CP-CH-LcyEi ( Figure 2-4 ).

[0029] 2. Creation of astaxanthin corn material

[0030] Maize embryos (1-1.5mm long) about 10 days after hybrid HiII maize pollination were used as the starting material, and maize transformation was carried out by Agrobacterium-mediated methods. The transformation process was as follows:

[0031] (1) Take Hi II young ears 10 days after pollination, first prepare 5% sodium hypochlorite so...

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Abstract

The invention discloses a combined expression vector and an application of the combined expression vector in corn kernel astaxanthin expression. The expression vector is prepared from pBDEN-CP-BZ, pBDEN-CP-BZ-LcyEi and pBDEN-CP-CH-LcyEi. According to the invention, functional genes and promoters carried in the vector are transferred into corn; astaxanthin in bred corn kernels reaches up to 100 micrograms per gram of dry weight of the corn kernels, which is about 7 times that of the corn kernels reported originally, and astaxanthin is mainly trans-astaxanthin with high biological activity, so that the new corn germplasm rich in high-quality trans-astaxanthin opens up a new way for filling up the astaxanthin demand gap in China.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an astaxanthin expression vector and its application, and a method for determining the optimum storage condition and biological utilization of astaxanthin in corn. Background technique [0002] Astaxanthin is a carotenoid that exists in various algae, microorganisms, and aquatic animals, and has rarely been reported in higher plants (Cunningham and Gantt, 2011). Due to its excellent antioxidant capacity and other biological activities, astaxanthin is widely used as a nutritional supplement in food, feed, health care products, pharmaceuticals and cosmetics and other products. In the feed industry, astaxanthin can be used as a feed additive for abalone, sturgeon, salmon, rainbow trout, red sea bream, crustaceans and ornamental fish, various poultry and pigs. According to in vivo and in vitro test results, astaxanthin has potential positive effects on cancer, hypertension, ...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12N15/82C12N15/29A01H5/10A01H6/46
CPCC07K14/415C12N15/8234C12N15/8257
Inventor柳小庆陈茹梅李素贞杨文竹周晓今
Owner捷康生物科技(海南)有限公司