Promoter bashing
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Promoter bashing is a technique used in molecular biology to identify the importance of a specific promoter for the proper expression of the gene. Specific point mutations are made and the transcription of the gene is then measured. The importance of a promoter can be observed by the level of transcription.
1. Clone the region of DNA thought to act as a promoter
2. Sequence the region
3. Digest with appropriate restriction enzymes
4. Ligate to reporter gene so that the reporter can be transcribed in the cells of interest
5. Transform cells of interest with the various promoter:reporter constructs
6. Measure reporter-gene transcription rates by assaying the reporter gene product
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