I have 5 sequenced genes, including the punitive promotor region of each gene. All are involved in the cold acclimation process in arabidopsis. Each is expressed only when the temperature falls below a certain point. the specific transcripition factor
CBF1 binds somewhere between -200 and -250 of the genes to initiate transcription.
How do i use the sequences to determine the sequence that is bound by CBF1?© BrainMass Inc. brainmass.com June 3, 2020, 6:23 pm ad1c9bdddf
There are two main approaches that you can use for determining the binding sequence for CBF1. Assuming that you have the purified protein, you can do a DNA fingerprinting technique. In this approach, you incubate the cloned and 32P-labelled DNA sequences with the purified CBF1 protein. After a period of time, partially digest the DNA with nucleases. Then, run the DNA fragments on a polyacrylamide gel using electrophoresis. Expose the gel to autoradiographic film. This will allow the 32P-labelled ...
The expert examines the DNA sequence motif that is bound by CBF1.