Retrovirus-like vectors for Saccharomyces cerevisiae: integration of foreign genes controlled by efficient promoters into yeast chromosomal DNA.
Using modified Saccharomyces cerevisiae Ty1 elements located on a 2 mu plasmid, reverse-transcriptase-mediated transposition into yeast chromosomes of expression cassettes containing a foreign gene can be induced. These expression cassettes consist of the yeast ARG3 and CUP1 promoter sequences fused to the Escherichia coli galK structural gene. Expression cassettes as large as 2 kb can be inserted into Ty elements and transposed efficiently to various sites in the yeast genome. A third yeast promoter (from the yeast CAR1 gene) seems to be unsuitable for use in the expression cassette. This may be because it does not allow the transcription run-through necessary for Ty1 transposition. Ways of improving this vector system are discussed, as are its advantages over episomal vector systems.
Jacobs
E
E
Molecular and Cellular Biology Department, Smith Kline-RIT, Rixensart, Belgium.
Dewerchin
M
M
Boeke
J D
JD
D001483:Base Sequence; D015139:Blotting, Southern; D002875:Chromosomes; D003470:Culture Media; D004247:DNA / Q000235:genetics*; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005686:Galactokinase / Q000032:analysis / Q000235:genetics; D005786:Gene Expression Regulation; D005796:Genes; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D003470:Culture Media; D004251:DNA Transposable Elements; D004247:DNA; D005686:Galactokinase