Getting the reporter gene in cells - Stable transformation - Different integration sites: Position Effect

5 important questions on Getting the reporter gene in cells - Stable transformation - Different integration sites: Position Effect

This position effect can be fixed by...

Flanking the reporter gene by MAR elements (matrix attachment region).

How can reaching the right position also be achieved?

By homologous recombination, so using a construct with flanking sides that are the same to the target gene.

What bacteria and how can it be used to also transform plants?

Agrobacterium rhizogenes; this one transforms with Ri-DNA, containing auxin synthesis genes inducing a hairy root culture.
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Name 3 advantages of using Agrobacterium rhizogenes.

1. Faster growth and scale-up capability
2. Grow without light -> no pigment -> no auto-fluorescence
3. Stable metabolite production

Name a third method to transform plants?

CRISPR-Cas9; bacterial antivirus mechanism discovered in E. Coli.
- Can be delivered into plant cells by Agrobacterium tumefaciens.

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