Gene for encoding GA20ox-oxidase and application of gene

A technology of oxidase and gene, applied in the field of genetic engineering, can solve the problems of reducing plant height and reducing endogenous GA content, and achieves the effect of simple and efficient method

CN112251451AActive Publication Date: 2021-01-22GUANGXI SUBTROPICAL CROPS RES INST GUANGXI SUBTROPICAL AGRI PROD PROCESSING RES INST
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2021-01-22

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Abstract

The invention belongs to the technical field of gene engineering, and particularly relates to a gene for encoding GA20-oxoxidase and application of the gene. The gene for encoding the GA20-oxoxidase is obtained from a constructed saccharomycetes cDNA library, and the nucleotide sequence of the gene is as shown in SEQ ID No. 1. According to the gene, a GA20ox gene sequence is directly obtained froma test material, the physicochemical activity of the gene is analyzed, and the gene is used as an index for regulating and controlling dwarfing traits. The invention relates to cloning of the GA20oxoxidase gene, analysis of an amino acid sequence corresponding to the GA20ox oxidase gene, and determination of the expression quantity of the GA20ox gene through a semi-fluorescent quantitative method, and provides a new method for early pre-judging whether a test material has dwarfing traits or not. The method for determining the expression quantity of the GA20ox gene through the semi-fluorescence quantitative analysis method is simple and efficient for early pre-judging whether the test material has the dwarfing characteristic or not.
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Description

technical field

[0001] The invention belongs to the technical field of genetic engineering, in particular to a gene encoding GA20ox-oxidase and its application. Background technique

[0002] Dwarf planting is a constant development trend of crop production. Plant dwarfing is mainly divided into genetic dwarfing and physiological dwarfing. A single gene mutation belongs to hereditary dwarfing, and its dwarfing incentive is mostly gene synthesis protein, which affects hormones, and hormones affect phenotype; the influence of hand and environmental factors leads to changes in plant phenotype, which belongs to physiological dwarfing, and its dwarfing effect The mechanism is mostly that environmental changes affect the secretion of plant hormones, which eventually lead to changes. But no matter what kind of dwarfing mode, the phenotype must be affected by hormones, and the hormones in the process of plant growth are mostly related to gibberellin.

[0003] The study of dwarf ge...

Examples

Embodiment

[0024] 1. Carrier and competent state

[0025] The cloning vector pGE-T Vector was purchased from Huada Gene Biotechnology (Shenzhen) Co., Ltd.; JM109 competence was preserved in the Key Laboratory of New Technologies for Breeding and Comprehensive Development of Superior Crops, Guangxi Tropical Crops Research Institute; subcellular localization vectors pK7FWGF2.0 and PEASY -T1 was provided by Jiakang Chuangyin Biotechnology (Wuhan) Co., Ltd.

[0026] 2. Main reagents

[0027] DNA Marker, T 4 DNA ligase, Ultra-fidelity DNA polymerase, 2×Taq Master Mix, FirstChoice RLM-RACE Kit, 5×RT Reaction Buffer, dNTP, RNaes Inhibitor, M-MLV RTase, gel recovery kit were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. The adapters and primers were synthesized by Huada Gene Biotechnology (Shenzhen) Co., Ltd.

[0028] 3. Method