Transfer of genes for utilization of starch (sta2) and melibiose (mel) to industrial strains of Saccharomyces cerevisiae by single-chromosome transfer, using a kar1 mutant as vector.
A method has been developed for the transfer of genes from other yeast strains and species to industrial yeast strains, using a haploid, kar1-1 mutant strain of Saccharomyces cerevisiae as a vector. The sta2 gene, conferring the ability to metabolize starch was transferred from an auxotrophic haploid strain of S. cerevisiae (S. diastaticus) and the melibiose-metabolism (mel) gene(s), from S. kluyveri, to the kar1-1 mutant [K5-5A; (alpha ade2 his4 can1 gal) by normal mating and protoplast fusion. From this strain, the genes were transferred to baker's yeast and brewing yeast strains, which did not utilize starch, and to baker's yeast strains, which did not utilize melibiose, by protoplast fusion, spore-cell pairing, or rare-mating. Strains that utilized starch or melibiose were obtained by all three methods. Pulsed-field gel electrophoresis preparations showed little change in the mobility of the chromosomes of the hybrids. The most probable explanation for the results obtained is that single chromosomes were transferred, first, from the donor strains to the kar1-1 haploid mutant strain, and then from the kar1-1 vector to the recipient industrial strain of S. cerevisiae. The transfer of the genes is probably accomplished through formation of disomic strains and then, in the case of the hybrids that metabolize starch, by integration of the sta2 gene into the genome of the industrial yeast strains.
Planta Piloto de Procesos Industriales Microbiológicos (PROIMI), Tucuman, Argentina.
J F
JF
Spencer
D M
DM
Spencer
L
L
de Figueroa
J M
JM
Nougues
H
H
Heluane
D015825:Chromosomes, Fungal; D004586:Electrophoresis; D005656:Fungal Proteins / Q000235:genetics*; D005696:Galactosidases / Q000235:genetics*; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics*; D008553:Melibiose / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D013172:Spores, Fungal / Q000235:genetics; D013213:Starch / Q000378:metabolism; D014162:Transfection / Q000235:genetics*
D005656:Fungal Proteins; C074559:KAR1 protein, S cerevisiae; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D013213:Starch; D008553:Melibiose; D005696:Galactosidases; D005087:Glucan 1,4-alpha-Glucosidase