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US 8,507,758 B2Granted
Markerless transformation
AssigneeSeminis Vegetable Seeds, Inc.
InventorJoseph Michael Palys
Life of this patent
Estimating the term…
Abstract
Methods for identification of successful transformation without a need for in vitro selection are provided. Direct detection of nucleotide sequences of interest is described which eliminate the need for use of ancillary nucleotide sequences.
Claims18
A method for producing a transgenic plant comprising a nucleotide sequence of interest inserted into its genome, where said method consists essentially of:
- a) transforming a plant tissue or plant cell with said nucleotide sequence of interest, which nucleotide sequence of interest comprises a gene of interest;
- wherein said plant tissue or plant cell is not transformed with an ancillary nucleotide sequence comprising a reporter sequence or a selectable marker sequence;
- b) regenerating said plant tissue or plant cell without selection for a reporter or selectable marker following said transforming, to produce a transgenic plant;
- and
- c) directly detecting the presence or absence in said transgenic plant of said nucleotide sequence of interest, an encoded product of said nucleotide sequence of interest, or a metabolic product of said encoded product;
- wherein said transgenic plant comprises said nucleotide sequence of interest introduced by said transforming;
- wherein said transforming does not introduce a transposable element into said plant tissue or plant cell;
- and
- wherein said nucleotide sequence of interest encodes one or more products selected from the group consisting of a hybrid Cry1Ac-1Acf Bacillus thuringiensis (Bt) protein, a Cry1Bb Bt protein, and a Cry1Ac Bt protein.
6 more elements
A method for producing a transgenic plant comprising a nucleotide sequence of interest inserted into its genome, where said method comprises:
- a) transforming a plant tissue or plant cell with said nucleotide sequence of interest, which nucleotide sequence of interest comprises a gene of interest;
- wherein said plant tissue or plant cell is not transformed with an ancillary nucleotide sequence comprising a reporter sequence or a selectable marker sequence;
- b) regenerating said plant tissue or plant cell without screening for said nucleotide sequence of interest, an encoded product of said nucleotide sequence of interest, or a metabolic product of said encoded product;
- and without selection for a reporter or selectable marker following said transforming, to produce a transgenic plant;
- and
- c) directly detecting the presence or absence in said transgenic plant of said nucleotide sequence of interest, said encoded product of said nucleotide sequence of interest, or said metabolic product of said encoded product;
- wherein said transgenic plant comprises said nucleotide sequence of interest introduced by said transforming;
- and
- wherein said nucleotide sequence of interest encodes one or more products selected from the group consisting of a hybrid Cry1Ac-1Acf Bacillus thuringiensis (Bt) protein, a Cry1Bb Bt protein, and a Cry1Ac Bt protein.
6 more elements
Dependencies are inferred from the claim text.