Integration of heterologous genes into the chromosome of Saccharomyces cerevisiae using a delta sequence of yeast retrotransposon Ty.
Distribution of a delta (delta) sequence of the Ty element on a chromosome of the yeast Saccharomyces cerevisiae was analysed by pulsed-field gel electrophoresis. More than 100 copies of the delta sequence were nonrandomly distributed on the chromosome. Using the delta sequence as a recombination site, mouse alpha-amylase and human beta-endorphin genes were introduced into the chromosomal DNA. The integration occurred on a particular chromosome in each case and the copy number was estimated as three to five. It was also found that single- or multi-copy integration occurred at a single or multiple sites on the particular chromosome. The integrants secreted alpha-amylase and beta-endorphin by three-to fivefold compared with single-copy integrants. This type of integration was mitotically stable over a period of 50 generations under non-selective conditions.
Laboratory of Molecular Genetics, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
A
A
Sakai
Y
Y
Shimizu
F
F
Hishinuma
D002875:Chromosomes / Q000378:metabolism*; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005800:Genes, Fungal; D010957:Plasmids; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D000516:alpha-Amylases / Q000235:genetics; D001615:beta-Endorphin / Q000235:genetics
D004251:DNA Transposable Elements; D001615:beta-Endorphin; D000516:alpha-Amylases