Selective fitness of four episomal shuttle-vectors carrying HIS3, LEU2, TRP1, and URA3 selectable markers in Saccharomyces cerevisiae.
A comparison of the selective fitness of four 2-microm-based shuttle-plasmids carrying the yeast genes HIS3, LEU2, TRP1, and URA3 was performed. The effect of each marker on long-term growth rate and plasmid maintenance was measured. In selective medium, the LEU2 and URA3 plasmids were maintained at the lowest and the highest levels, respectively, while the HIS3 and TRP1 plasmids were maintained at an intermediate level. In synthetic complete medium, plasmid loss rate was lower for the genes TRP1 and URA3 than for the other two markers, and a similar pattern was observed for cells growing in rich medium. These results were confirmed by competition experiments among transformants with different plasmids in complete and rich media, indicating a different degree of fitness for the markers used. A potential correlation of the energy cost of plasmid maintenance with the secondary DNA structure and the level of expression of the selective markers is also investigated.
Ugolini
Simone
S
Microbiology Group, International Centre for Genetic Engineering and Biotechnology, AREA Science Park, Padriciano 99, I-34012 Trieste, Italy.
Tosato
Valentina
V
Bruschi
Carlo V
CV
D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D019747:Aldose-Ketose Isomerases; D002455:Cell Division / Q000235:genetics; D003470:Culture Media; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics*; D006836:Hydro-Lyases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D012641:Selection, Genetic; D014158:Transcription, Genetic
D003470:Culture Media; D005656:Fungal Proteins; D005819:Genetic Markers; D029701:Saccharomyces cerevisiae Proteins; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase; D019747:Aldose-Ketose Isomerases; C490353:TRP1 protein, S cerevisiae