Vacuum Infiltration of seedling plants
Patent application filed by The Samuel Roberts Noble Foundation
Actual pending claims
EP 1141356 A2 |
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Claim 1A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with Agrobacterium cells, said Agrobacterium cells harboring a vector, said vector enabling said Agrobacterium cells to transfer T-DNA containing at least one gene or gene fragment to said seedling; B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer said T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different. |
Claim 9A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with a mixture of Agrobacterium cells, said mixture comprising cells from a Agrobacterium strain harboring a vector with a DNA fragment and cells from said Agrobacterium strain harboring said vector a second DNA fragment, said vector enabling said Agrobacterium cells to transfer said T-DNA to said seedling; B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different. |
Claim 18A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with Agrobacterium cells, said Agrobacterium cells harboring a vector, said vector enabling said Agrobacterium cells to transfer T-DNA containing at least one gene or gene fragment and a selectable marker gene to said seedling; B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer said T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different; C) allowing said transformed seedling to grow to maturity and set seed; D) germinating said seed to form progeny; E) exposing said progeny to an agent enabling detection of selectable marker gene expression; F) selecting for progeny expressing said selectable marker gene and at least one gene, said expression of said selectable marker gene and at least one gene indicating gene transfer. |