Discussion
Which of the following statements are TRUE on transgene approach?
P. T-DNA integration occurs mainly through non-homologous recombination
Q. The Gateway cloning depends on recombination technology as opposed to standard uses of restriction enzymes and DNA ligase
R. The localization of β-glucuronidase (GUS) activity as a result of expression of GUS reporter gene can be visualized in a histochemical assay using the X-gal
S. The green fluorescent protein gene (GFP) is isolated from the bacterium Photinus pyralis
Answer: A
The true statements are P, Q.
P is true: Agrobacterium T-DNA integrates into the plant genome mainly by illegitimate (non-homologous) recombination, at essentially random sites. Q is true: Gateway cloning moves inserts between vectors by site-specific recombination at att sites, using the phage lambda integrase system rather than restriction enzymes and ligase.
R is false: GUS activity is detected histochemically with X-Gluc (5-bromo-4-chloro-3-indolyl glucuronide); X-gal is the substrate for β-galactosidase. S is false: GFP comes from the jellyfish Aequorea victoria; Photinus pyralis is the firefly that supplies the luciferase gene.
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