Methods and compositions to improve the spread of chemical signals in plants

A compound and chemical technology, applied in the field of molecular biology, can solve the problem of limiting the degree to which chemical gene switches can be activated

Inactive Publication Date: 2016-04-06
PIONEER HI BRED INT INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This limits the extent to which chemical gene switches can be activated in tissues or organisms that cannot readily come into contact with chemical ligands

Method used

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  • Methods and compositions to improve the spread of chemical signals in plants
  • Methods and compositions to improve the spread of chemical signals in plants
  • Methods and compositions to improve the spread of chemical signals in plants

Examples

Experimental program
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Effect test

example 1

[0319] Example 1. Use of siRNAs controlled by sulfonylurea repressors targeting sulfonylurea repressors to amplify and enhance induced The spatial distribution of the pilot signal

[0320] One of the major limitations of any chemical induction system in multicellular organisms is the penetration and uniform distribution of the inducer in all tissues (due to altered motility or metabolism). The consequence is that induction of the target gene may be uneven (or absent) in the tissue or cell type of interest. To address this potential problem, it is desirable to provide additional genetic factors to influence the spread of derepression. siRNA has been widely used in eukaryotic systems to reduce target gene expression. In particular, plants have an increased likelihood that the siRNA response can proceed systemically (Palauqui et al. (1997) EMBO J. 16:4738-4745 (Palauqui et al., 1997, "European Molecular Biology Organization Journal" Vol. 16 No. 4738 -4745 pages); Voinnet et...

example 2

[0329] Example 2. Use of miRNAs controlled by sulfonylurea repressors targeting sulfonylurea repressors to amplify and enhance induced The spatial distribution of the pilot signal .

[0330] To show that the presence of an amiRNA targeting a repressor increases expression after induction, two constructs were made. The first construct, the control construct pPHP46916 (10,904bp) (SEQ ID NO: 2111 ), contained the cassette in which the soybean S-adenosylmethionine promoter was operably linked to the soybean acetolactate synthase gene with the HrA mutation, with The HrA mutated soybean acetolactate synthase gene is operably linked to cassette A of the soybean acetolactate synthase terminator (this cassette is used as a selectable marker during plant transformation; positions 81-4062); followed by the T7 promoter in which To the hygromycin phosphotransferase gene, the hygromycin phosphotransferase gene is operably linked to cassette B of the T7 terminator (this cassette is used ...

example 3

[0337] Example 3 Preparation of Soybean Embryo Culture and Model System Transformation Using Soybean Expression Vectors and Implantation Strain regeneration

[0338] Culture conditions :

[0339] Soybean embryogenic suspension cultures (cv. Jack) were maintained in 35 mL of liquid medium SB196 (below) on a rotary shaker at 150 rpm at 26 °C with cool white fluorescent lights and a photoperiod of 16:8 hours Day / night, the light intensity is 60-85μE / m2 / s. Cultures were subcultured every 7 days to two weeks by inoculating approximately 35 mg of tissue into 35 ml of fresh liquid SB196 (the preferred subculture interval was every 7 days).

[0340] Soybean embryogenic suspension cultures were transformed using soybean expression plasmids by the particle gun bombardment method (Klein et al., Nature 327:70 (1987) (Klein et al., "Nature" Vol. A HE instrument (helium modification) performed all transformations.

[0341] Priming of soybean embryogenic suspension cultures :

[...

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Abstract

Compositions and methods are provided which employ a chemical-gene switch. The chemical-gene switch disclosed herein comprises at least three components. The first component comprises a polynucleotide encoding a chemically-regulated transcriptional repressor; the second component comprises a repressible promoter operably linked to a polynucleotide of interest, and the third component comprises a gene silencing construct operably linked to a second repressible promoter, wherein the gene silencing construct encodes a silencing element that decreases the level of mRNA encoding the chemically -regulated transcriptional repressor. Expression of the polynucleotide of interest and the silencing construct is controlled by providing the appropriate chemical ligand. Transient induction from the chemical ligand leads to the production of the silencing element, and the destruction of the mRNA encoding the chemically-regulated transcriptional repressor. The presence of the silencing element maintains a state of de-repression. In specific embodiments, the silencing elements are cell non-autonomous, the state of de-repression becomes more distributed throughout the plant beyond where the chemical ligand reaches.

Description

[0001] References to Sequence Listings [0002] The Sequence Listing is a text file named 36446_0068P1_Seq_List.txt, created on March 5, 2014, filed on March 11, 2014, and is 2,258,550 bytes in size, hereby pursuant to Title 37 of the US Patent Regulations (C.F.R.) Section 1.52(e)(5) is hereby incorporated by reference. technical field [0003] The present invention relates to the field of molecular biology, and more particularly, to the regulation of gene expression. Background technique [0004] The tetracycline operator system, comprising repressor and operator elements, was originally isolated from bacteria. This operon system is tightly controlled by the presence of tetracycline and itself regulates the expression levels of the tetA and tetR genes. The product of tetA removes tetracycline from the cell. The product of tetR is the repressor protein bound to the operator element, which in the absence of tetracycline K d at about 10 pM, thereby blocking the expression ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC12N15/8217C12N15/8238C12N15/8218
Inventor K.E.麦克布里德B.麦克戈尼格尔N.S.亚达夫
Owner PIONEER HI BRED INT INC
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