Construction of promoter-probe vectors for Candida maltosa,a n-alkane-assimilating yeast,using the LEU2 gene of Saccharomyces cerevisiae

For the purpose of isolation of promoter regions which are regulated by a carbon source in the medium in an n-alkane-assimilating yeast, Candida maltosa, two promoter-probe vectors were constructed. Each of them consists of the LEU2 gene of Saccharomyces cerevisiae whose 5′-non-coding region was trimmed with BAL31, an autonomously replicating sequence isolated from C. maltosa genome (the TRA region) which we have previously isolated, and the pBR322 sequence. One of them, pPLC2, having the TATA box, lacks the regulatory sequence (“sequence L”) of the LEU2 gene, and the other, pPLC1, lacks both the TATA box and sequence L. Using pPLC1 as a shot-gun cloning vector in C. maltosa, many promoter regions which were active when glucose was present in the medium as a carbon source were obtained from the genome of C. maltosa. The sizes of the inserted fragments of two of them were determined. (In this paper, a promoter region refers to a promoter which includes a TATA box, plus a regulatory sequence such as an UAS (upstream activating sequence)-like sequence).