Citrus cell capable of generating beta-citraurinene for polygenic metabolism engineering building and application
A cell and citrus technology, applied in the field of citrus callus cells, can solve the problems of unreported plant callus cells, restricted substance research and application, and rare substances, so as to get rid of the influence of external natural environmental conditions, solve supply cycle restrictions, shorten the effect of time
Active Publication Date: 2018-08-24
HUAZHONG AGRI UNIV
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Problems solved by technology
The first two substances have already been produced by companies as standard products, but for the third substance β-citraurinene (molecular formula is ), currently only one of our patents (Application No. 201710807932.0) discloses an HPLC method for separation and purification, and there is no literature disclosing plant callus cells that can directly produce β-citraurinene, nor is there any report on the construction of β-citraurinene by genetic manipulation Plant callus cells that can produce β-citraurinene make this substance extremely rare, limiting the research and application of this substance
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[0024] The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.
[0025] 1. Basic Experimental Procedures
[0026] 1.1 Preparation of Agrobacterium infection solution
[0027] 1) Agrobacterium monoclonal activation: Inoculate the detected Agrobacterium monoclonal liquid carrying the target gene into solid LB medium (spec final concentration 100mg / L plus a small amount of rifampicin), in a constant temperature incubator at 28°C for 2 days;
[0028] 2) Propagation of Agrobacterium: Pick a single clone for further verification of positive results, then use an inoculation loop to pick up the positive single clone of Agrobacterium, and streak (a little denser) on a solid LB plate (containing spec 100mg / L without adding rifampicin) , 28°C constant temperature incubator for 2 days;
[0029] 3) Prepare the Agrobacterium ...
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The invention relates to a citrus callus cell capable of generating beta-citraurinene. The cell belongs to the citrus callus cell with a CrtB gene super expression frame and a CCD4b gene super expression frame. The invention also relates to application of the citrus callus cell to beta-citraurinene production, and also relates to a method for producing beta-citraurinene. The method comprises the step of culturing the citrus callus cell and extracting the beta-citraurinene. By using the citrus callus cell and the method provided by the invention, the beta-citraurinene can be produced through callus cell culture; the problem of supply period limitation (the plant extraction can only be performed after the fruit mature in December) is solved; the influence of the external natural environmentconditions is avoided; in addition, the types of carotenoids in the callus cells are few; after the mass generation of the substances in the callus tissues, the purification is facilitated; the advantages are favorable for mass production of the C30 rare carotenoid; the method is suitable for biological factory production of the substances.
Description
technical field [0001] The invention relates to the field of tissue metabolism engineering, more particularly, relates to a citrus callus cell capable of producing β-citraurinene and application thereof. Background technique [0002] Carotenoids are an important natural pigment that exists widely in nature, and are a class of C pigments composed of isoprene skeletons. 40 or C 30 of terpenoids. Carotenoid molecules appear yellow, orange or red, embellishing a colorful world for human life. With the in-depth study of carotenoids, more and more studies have shown that carotenoids and their derivatives are important health care products for human nutrition and health, and play an important role in human health and food processing. Some carotenoid components have the effects of improving human immunity, reducing the risk of cardiovascular diseases, preventing chronic diseases and cancer, and ensuring human health and reproductive development; carotenoids are also important nat...
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IPC IPC(8): C12N5/10C12P23/00
CPCC12N5/04C12P23/00
Inventor 邓秀新郑雄杰朱凯杰汤雨晴叶俊丽曹洪波
Owner HUAZHONG AGRI UNIV