Question

Using your knowledge of reverse genetics, devise an experiment that uses transgenic plants to demonstrate that proteins can move from the cells in which they are produced to other cells in the plant.

Answer 1

Forward genetics = From phenotype to genotype
Reverse genetics = From genotype to phenotype

In reverse genetics, we will study the phenotype of a known mutant.

Cell-autonomous function: The protein function is observed in the cell in which it is produced
Non-cell-autonomous function: The protein function is observed in other cells in which it is not produced i.e. the protein moves

To demonstrate that a protein is moving, clone the gene of interest under its own promoter along with a heavy tag such as 3X GFP or 2X MBP. These heavy tags prevent protein mobilization.
Transform this construct into the mutant of the gene of interest and create a transgenic.
Observe the GFP signal. It should be seen only in the cell it is expressed (We can compare promoter reporter and protein reporter lines).

If promoter reporter lines and protein reporter lines exhibit the same expression pattern, it means that the protein is not moving.
If protein reporter line exhibits an expanded expression as compared to the promoter reporter line, it shows that the protein is moving.