Functional Analysis of the Upstream
Sequence of PHO81
Gene of Saccharomyces cerevisiae
XIA Zan-Xian and AO Shi-Zhou*
( State Key Laboratory of Molecular Biology, Shanghai Institute of
Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China
)
Abstract Deletion analysis on the fused
PHO81-lacZ gene revealed two important regions in the upstream sequences of
PHO81 gene：
－401―－289 bp and －1 012―－801 bp. They did not share
higher similarity with the upstream regions of PHO5 and PHO84
gene, except that －401―－289 bp contains the 5′-CACGTG/T-3′
motif, which was found among the upstream regions of PHO5 and PHO84
gene and was the core sequence of PHO4 binding site； it also contains
A/T-rich segments flanking the motif, which may be PHO2 binding sites. This
suggests that the －401―－289 bp of the upstream region
of PHO81 gene may contain upstream activation sequence (UAS), and －1012―－801 bp may contain upstream
enhancer sequence. The gel retardation assays of the －1 012―－801 bp was performed using
yeast total protein extract, and the results showed that there was an unknown
protein factor binding at the region.
Key Words PHO81 gene; deletion analysis; retardation
assay
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