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How would you experimentally identify which subset of the sequences in the 7-kb fragment are required...

How would you experimentally identify which subset of the sequences in the 7-kb fragment are required for the normal expression of wg?

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Answer #1

Deletion mapping:

It is a molecular biology method in which fragments of upstream regulatory sequences are fused with a reporter gene such as GFP or GUS and transformed. If the given cis element is sufficient to cause expression of the reporter gene, we can observe the GFP or GUS signal.

So, clone different fragments of varying lengths of the upstream 7 kb sequence into different constructs containing GFP. Whichever fragment containing clone expresses GFP signal, that is most likely responsible for the expression of the gene in vivo.

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