Wheat 3-hydroxy-3-methylglutaryl-CoA reductase gene tahmgr and its isolation and clone, site-directed mutagenesis and detection method of enzyme function

Abstract

The invention belongs to the field of molecular biology, and relates to an isolation and cloning, gene site-specific mutagenesis, zymoprotein prokaryotic expression, zymoprotein separation and purification, and enzymatic activity detection technology for a first key enzyme gene participating in isoprenoid substance synthesis in wheat mevalonic acid metabolic pathways. Important technical reserves are provided for further carrying out gene modification and augmentation, constructing eukaryotic genetic expression vectors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) genes and converting corresponding products, and are particularly provided for obtaining plants with the high commercial value through secondary metabolism, discussing the relationship between overexpression of the HMGR genes in the receiver plants and important economical characters such as secondary metabolism products, the crop grain size and the grain weight, then increasing the crop yield, analyzing the structures of introns, exons and promoters of the HMGR genes, researching the functions of the promoters and developing related molecular markers.

Description

technical field

[0001] The invention belongs to the field of molecular biology and relates to wheat 3-hydroxy-3-methylglutaryl-CoA reductase, the first key enzyme gene involved in the synthesis of isoprenoid substances in the metabolic pathway of wheat mevalonate Gene TaHMGR and its isolation and clone, site-directed mutation and detection method of enzyme function. Background technique

[0002] Isoprenoid substances are important substances necessary for maintaining plant growth and development, photosynthesis, electron transfer, and coping with environmental stress. Such substances can be used as photosynthetic pigments (such as chlorophyll, carotenoids), growth substances and plant hormones (such as cytokinins, abscisic acid, gibberellins and brassinosteroids), part of membrane structures such as sitosterol, electron transport Receptors such as plastoquinone, receptors for glucose in glucosylation reactions such as dolichol, and can regulate cell growth (such as prenyl p...

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