Analysis of selective gene activation in yeast by differential display

TATA box binding protein-associated factors (TAF(II)s) are dispensable for transcription of most genes in yeast. To further investigate the in vivo functions of TAF(II)s, differential display was used to identify a small subset of yeast genes whose transcription is dependent on yeast TAF(II)145, the core TAF(II) component that contacts TATA box-binding protein (TBP). Messenger RNA profiles derived from a wild-type TAF(II)145 strain and a temperature-sensitive taf(II)1145 strain were analyzed. Those genes whose messenger RNA level was greatly reduced in the temperature-sensitive taf(II)145 strain were cloned and further characterized. The procedure for performing differential display described here is modified from protocols provided by the manufacturer (Display Systems) and optimized for the yeast system.