A consensus transcription termination sequence in the promoter region is necessary for efficient gene expression of the TRP1 gene of Saccharomyces cerevisiae

Abstract

The TRP1 gene of Saccharomyces cerevisiae is the only TRP gene which is not derepressible by the general control regulatory system. In the TRP1 promoter transcription starts at five initiation sites, organized in two clusters. The two transcripts of the first, more upstream cluster include a long leader sequence of approximately 200 bp. A transcriptional terminator element located in the 5' region of the TRP1 gene is essential for accurate gene expression. In partial TRP1 promoters lacking the terminator, like the original EcoRI TRP1 fragment used in numerous vectors, plasmid-encoded transcription is initiated predominantly in adjacent vector regions, resulting mainly in large, poorly translated transcripts. This poor translation is not due to mRNA instability. The effect can be suppressed by introducing artificial transcription barriers between vector sequences and the truncated EcoRI TRP1 fragment.