Lab 4 - brassica transformation and arabidopsis screening Flashcards
1
Q
cocultivation
A
explant dipped in agrobacterium. it infects the cells of cut surface. The cells are placed in high BA medium with no antibiotics. BA promotes cell division
2
Q
selection
A
after kill stage, T-DNA inserted into cell by agrobacterium contains a selectable marker gene (NPT II) is placed on media with kanamycin (which selects for cells with T-DNA)
3
Q
antibiotic use
A
kanamycin will kill off cells that are not transformed
4
Q
why does petiole transformation work
A
5
Q
regeneration and selection
A
regeneration - cell subject to hormones and is induced to become entire plant (formation of organs)
selection - selectable marker gene used, kanamycin is able to select for it
6
Q
selectable markers
A
- encode traits that are easily detectable
7
Q
screening
A
- sterilize seed from transgenic plant and test seed for presence of selectable marker gene
