Molecular Cloning and DNA Analysis of the Orotidine-5′-Phosphate Decarboxylase Gene from the Yeast Saccharomyces exiguus Yp74L-3

Abstract

The orotidine-5'-phosphate decarboxylase gene of Saccharomyces exiguus Yp74L-3 was cloned as a DNA fragment complementing a ura4 mutation of this yeast. The coding region of the gene is 807 bp in length, and represents 68.7% similarity to the corresponding gene of S. cerevisiae (URA3). The cloned URA4 gene was shown to be located on the 790-kbp Chromosome (chr) VIII of S. exiguus Yp74L-3. The neighbor-joining phylogenetic tree based on the orotidine-5'-phosphate decarboxylase coding sequences indicates that S. exiguus Yp74L-3 is closely related to Kluyveromyces yeasts, as well as to a S. cerevisiae laboratory strain.

Molecular cloning and DNA analysis of a gene encoding α mating pheromone from the yeast Saccharomyces naganishii

“…Plasmid insert DNA containing the α mating pheromone gene was subjected to a series of deletions through digestion with various restriction endonucleases and recircularization (Sambrook and Russell, 2001). Southern hybridization was carried out using the α mating pheromone probe labelled with a non‐radioisotope tracer, as described in Hisatomi et al (2000).…”

“…DNA sequencing was performed using an automated system according to Hisatomi et al (2000). A commercial software program, Genetyx‐Mac (Software Development, Tokyo, Japan), was used to search for an open reading frame and to calculate a hydrophobicity profile of a precursor peptide of α mating pheromone deduced from the DNA sequence.…”

“…Separation of chromosomal DNAs and chromosome assignment of a specific DNA fragment were performed using a CHEF apparatus and a non‐radioisotope hybridization technique according to Hisatomi et al (2000).…”

“…To shed light on the mechanism of speciation based on sexual isolation in the genus Saccharomyces , we previously studied sexual differentiation in Saccharomyces exiguus Yp74L‐3 in comparison with that of S. cerevisiae (Hisatomi et al , 1986, 1988, 1998, 1999, 2000; Hisatomi and Tsuboi, 1989). Recently, the phylogenetic position of S. exiguus Yp74L‐3 was reexamined.…”