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Method of excising a nucleic acid sequence from a plant genome

A nucleic acid sequence and genome technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, plant products, etc., can solve problems such as deletion and chromosome rearrangement

Inactive Publication Date: 2010-11-17
BASF PLANT SCI GMBH
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Problems solved by technology

May result in significant chromosomal rearrangements or deletions

Method used

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  • Method of excising a nucleic acid sequence from a plant genome
  • Method of excising a nucleic acid sequence from a plant genome
  • Method of excising a nucleic acid sequence from a plant genome

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[0282] Materials and General Methods

[0283] Unless otherwise stated, chemicals and reagents in the examples were obtained from Sigma Chemical Company (St. Louis, MO), restriction endonucleases were from New England Biolabs (Beverly, MA) or Roche (Indianapolis, IN), oligonucleases were obtained from Nucleotides were synthesized by MWG Biotech Inc. (High Point, NC), and other modifying enzymes or kits for biochemical and molecular bioassays were from Clontech (Palo Alto, CA), Pharmacia Biotech (Piscataway, NJ), Promega Corporation (Madison, WI) or Stratagene (La Jolla, CA). Materials for cell culture media were obtained from Gibco / BRL (Gaithersburg, MD) or DIFCO (Detroit, MI). Cloning steps performed for the purpose of the present invention such as restriction cutting, agarose gel electrophoresis, purification of DNA fragments, transfer of nucleic acids to nitrocellulose and nylon membranes, ligation of DNA fragments, transformation of E. coli cells, cultivation of bacteria, ...

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Abstract

The present invention relates to a method for excising a nucleic acid sequence from the genome of a plant or a plant cell. This method is based on the steps of transforming a plant cell with a construct encoding a DNA double strand break inducingenzyme (DSBI), generating a transgenic plant line, performing a transient assay to analyze the functionality of the transgenic enzyme, crossing the plant line with a line containing a nucleic acid sequence to be excised and performing an immature embryo conversion or a tissue culture regeneration through callus formation. The method can also be reversed, which means that a plant cell is transformed with a construct encoding a nucleic acid sequence to be excised, and the crossing is performed witha plant line containing a DSBI. As an alternative to the crossing step, a re-transformation of a transgenic plant line with a second construct can also be performed. The invention is also directed to a plant obtained by this method, or progeny, propagation material, part, tissue, cell or cell culture, derived from such a plant. Finally, the invention relates to the use of a plant or progeny, propagation material, part, tissue, cell or cell culture, derived from this method, as aliment, fodder or seeds or for the production of pharmaceuticals or chemicals.

Description

Summary of the invention [0001] The present invention relates to methods for excision of nucleic acid sequences from the genome of plants or plant cells. The method is based on the steps of transforming plant cells with a construct encoding a DNA double-strand break inducing enzyme (DSBI), generating a transgenic plant line, performing a transient assay to analyze the functionality of the transgenic enzyme, and combining the plant line with Lines of nucleic acid sequences are crossed and subjected to immature embryo transformation or tissue culture regeneration by callus formation. The method can also be performed in reverse, meaning that plant cells are transformed with the construct encoding the nucleic acid sequence to be excised, and crossed with a plant line containing the DSBI. As an alternative to this crossing step, retransformation of the transgenic plant line with the second construct can also be performed. The invention also relates to plants obtained by such meth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00C12N9/22
CPCC12N15/8201C12N9/22
Inventor H-S·宋F-M·赖C·E·罗切J·A·布朗
Owner BASF PLANT SCI GMBH
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