Biology Forums - Study Force

I think that I understand the principle correctly:

-> insert a promotorless DNA construct that consists of a    3' splice donor + reporter gene +5' polyadenylation site

But how could I make the construct jump into an intron?
For example into a plant genome?
Is it done by transposons somehow?

Question: why place it into an intron? Just wondering ???

I wish I could use my mother tongue to explain that...
But I'll have a try ;)

If you place the construct into an intron, there won't be an damaged exon.

Imagine the construct jumps into an intron. Then the original 5' splice-acceptor uses the 3' splice donor of the construct to
cut out the intron. Then the poly(a)-sites terminates the whole thing.
Afterwards you will have  some of the first original exons + reportergene. (= a fusion protein)

If the reporter gene is a GFP you can use it to detect the rate of transcription by light emission and if you are lucky you can
find out where the original protein is localised in the cell (e.g. as a transcription factor in the core or as a trans membrane protein)

Question: But wouldn't you require some kind of enhancer region to translate the gene? Otherwise, you would have to place this transgene into an operon.

Yes of course. You have to insert you construct into a intron of an open reading frame after a promotor.

An alternative method is called "enhancer trap". There you have a construct with a minimal promotor that can be inserted anywhere but near an enhancer regulated region.