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A method for inhibiting the expression of gibberellin synthetic gene in pear seeds

A seed gibberellin and synthetic gene technology, applied in the fields of developmental biology and cell biology, can solve the problems of easy protein decomposition, time-consuming and labor-consuming, complicated requirements, etc., and achieve easy operation, practical application, and simple method Effect

Active Publication Date: 2020-05-19
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these techniques are complex, time-consuming and labor-intensive, and the results obtained are not satisfactory.
When activating the suppressor gene of the target gene, the targeting is not accurate; when adding the protein that suppresses the target gene, the protein is easily decomposed and the effect is poor

Method used

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  • A method for inhibiting the expression of gibberellin synthetic gene in pear seeds
  • A method for inhibiting the expression of gibberellin synthetic gene in pear seeds
  • A method for inhibiting the expression of gibberellin synthetic gene in pear seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This example takes the pear seed as the experimental material, and the process is as follows ( figure 1 ):

[0044] 1. Design of complementary chain GA20-ODN3:

[0045] According to the gene sequence of Pyrus GA20-oxidase (accession number: HQ833589) provided by GenBank, input the full sequence of GA20-oxidase mRNA into the new sequence editing window of RNAdraw software, select the calculation structure, and set relevant parameters as required. The free energy minimum principle yields simulated secondary structures. According to the primary structure of the target mRNA and the simulated secondary structure, the original clasp loop, internal loop, expansion loop and multi-branched loop are selected, and the 18nt GA20ODN3 is designed according to the principle of complementary base pairing. GA20-ODN3 sequence: 5'-TCTGACACGGTGGGTAGT-3', synthesized by phosphorothioate modification;

[0046] 2. Seeds are prepared by using the water selection method to select full partic...

Embodiment 2、 example 3、 example 4

[0054] Embodiment 2, example 3, example 4, method is the same as example 1.

[0055] The designed complementary chains GA20-ODN1, GA20-ODN2, and GA20-ODN4 have been proved by experiments to inhibit the synthesis of GA20 oxidase, and the process will not be repeated here.

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Abstract

The invention discloses a method for inhibiting pear seed gibberellin synthetic gene expression, relates to design of complementary chains and cultivation of degreasing cotton wrapped seeds, and belongs to the field of developmental biology and cell biology. According to the method for inhibiting pear seed gibberellin synthetic gene expression by the complementary chains, the influence of lack of gibberellin on seed germination rate can be more effectively learned, and the function of the gibberellin on breaking seed dormancy is more effectively researched. The method is simple, convenient and easy to operate and practically apply and can be widely used for inhibiting other gene expression of different plant seeds.

Description

technical field [0001] The invention relates to a method for inhibiting the expression of gibberellin synthetic genes in seeds, relates to the design of complementary chains and the cultivation of seeds wrapped in absorbent cotton, and belongs to the fields of developmental biology and cell biology. Background technique [0002] Gibberellin (GA) can promote seed germination. GA20-oxidase is an important regulator and rate-limiting enzyme of GA biosynthesis. GA20-oxidase is a soluble dioxygenase, which can catalyze the three-step oxidation reaction from GA12 to GA9 and GA53 to GA20. It is a multi-gene regulation and is the most studied enzyme among the key enzymes of gibberellin synthesis. . [0003] At present, it has become an international research trend to use some technologies to regulate the specific expression of related genes. Gene expression is regulated in four stages: signal transmission, gene knockout, mRNA transcription, and protein translation. However, thes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82
CPCC12N15/113C12N15/8218C12N2310/11
Inventor 王春雷孙娅刘正阳张盼盼黄旭
Owner YANGZHOU UNIV
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