Abstract
To identify transcription factors (TFs) that would interact with 5' upstream region of a gene, yeast one-hybrid screening (Y1H) is often used. For Y1H, we need to identify cis-regulatory region and multimerize it for placing them in upstream of reporter gene. In addition, large-scale screening is required because non-TF genes mostly occupy cDNA library. To overcome such problems, we constructed novel cDNA library that only contains TF genes. Results of our test experiments showed that several transcription factors were identified with efficiency more than 100-fold even though whole 500-1000bp promoter region was directly used as a probe. However, false-positive or false-negative result was sometimes observed in the yeast experimental system. Therefore we are now trying to develop new method using plant instead of yeast. We will report the results of comparison of these methods to identify upstream transcription factors.
