How can I clone the promoter region of my gene
Posted 29 May 2002 - 11:22 PM
Posted 31 May 2002 - 04:24 PM
Posted 15 June 2002 - 06:55 AM
First you must be sure is the region really the promoter or might it be that there is an excisting not translated Exon. For this propose I would perform a 5`-RACE or Primer Extension. Next step: If you know the sequence order
primer, make a PCR and clone the fragment. If not the simplest wy is first to sequence the promoter region tnah make a PCR or subcloning.
If you are going to perform reporter assays I would start with a fragment up to 1500kb of the 5`-promoter region, but also construct shorter constucts
Posted 15 June 2002 - 06:47 PM
It seems it is not an easy job to get a gene's promoter region even there are publications shows where it is and what the sequence is .