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Bacterial plasmid pBR322 sequences serve as upstream activating sequences in Saccahromyces cerevisiae
Author(s) -
Sidhu Rajinder S.,
Bollon Arthur P.
Publication year - 1990
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320060307
Subject(s) - pbr322 , biology , plasmid , saccharomyces cerevisiae , gene , microbiology and biotechnology , yeast , genetics
The expression of acid phosphatase (Apase) from PHO5 and MF α‐ PHO5 hybrid genes is regulated by inorganic phosphate and mating type locus respectively, as well as the PHO4 and MAT α 1 gene products respectively. When PHO5 and MF α‐ PHO5 hybrid genes were cloned in the Bam HI site of the pBR322 sequence of the yeast shuttle vectors (YRp7 or YEp9T), in one orientation they were regulated normally but in the other orientation their expression was not regulated but expressed constitutively. The pBR322 sequences present upstream of the inserted genes are responsible for the constitutive expression. By replacing the PHO5 upstream activating sequences (UAS) element with pBR322 fragments, we have identified three pBR322 sequences, rom nucleotides 376 to 650, 2068 to 2116 and 2136 to 2247, which were able to promote expression of APase. A comparison of these three pBR322 fragments revealed 5′ ATCGCGCGAG 3′ and 5′ CGGTGATGNCGG 3′ to be the common sequences likely to act as USAs in Saccharomyces cerevisiae . By using synthetic oligonucleotides, it was found that both sequences are required for maximum expression of APase activity.