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ID 15266

Genetically engineering plants using agrobacterium, Robert Horsch

Description:
Interviewee: Robert Horsch. Robert Horsch talks about agrobacterium as a ready-made delivery system for getting foreign DNA into plants. (DNAi Location: Manipulation > Techniques > Transferring & storing > Interviews > Bacterial transfer)
Transcript:
Well once we had the engineered gene, the selectable marker, and had it put in a vector in the agrobacterium, the rest was the, the cell culture and selection. So we would start with a bleach-treated petunia leaf that the bleach kills the surface microorganisms that might contaminate things, and then we use actually a common paper punch that's been soaked in alcohol to clean it up, and punch discs out of the leaf. These discs then are nice uniform size pieces of the leaf tissue and have a wounded edge all the way around them. The agrobacterium grown in a simple broth then could be innoculated into that wounded edge and give a uniform infiltration of the bacterium with the wounded plant cells. After about two days of this co-cultivation, you could use an antibiotic that would kill the agrobacterium, because it had already transferred its DNA from itself into the plant genome. And then you could start selecting for the selectable marker and stimulating the regrowth of individual plant cells in the edge of the leaf disc to grow back to a whole plant.
Keywords:
selectable marker,plant genome,manipulation techniques,paper punch,plant transformation,plant cells,agrobacterium,leaf tissue,dnai,uniform size,size pieces,interviewee,cell culture,regrowth,bacterium,genetic engineering,microorganisms,infiltration,delivery system,bleach
Creative Commons License This work by Cold Spring Harbor Laboratory is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 United States License.

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