Preliminary Experiements on the ilv 1 Gene of Saccaromyces Cerevisiae, Located on a Plasmid Recombined from a Yeast Plasmid, An e. Coli Plasmid and an Insert of Yeast Chromosomal DNA
This report presents the results of three experiments on a recombinant plasmid called the ilv 1 plasmid. This plasmid carries the ilv 1 gene of Saccharomyces cerevisiae which produces Threonine deaminase. In the first experiment the presence of this plasmid in a strain of S. cerevisiae called 3971 (i1v 1) is proven by the observation that 50% of the cells in a given clone will segregate the ilv 1 plasmid. The second experiment involved the determination of the specific activity of Threonine deaminase in strain 3971 (i1 v 1) and a wild type strain, CBII, after growing the cells in repressing (+I1e) and non-repressing (-lIe) media. The results show that the basal level of Threonine daminase in 3971 (i1v 1) is four-fold that of the control, CBll. The specific activity of Threonine dearninase in 3971 (i1v 1) cultured non-repressing medium is I5-fold the specific activity of Threonine daminase in repressing medium. The result however does not prove that the ilv 1 gene is being regulated by lIe because only 2% of the 3971 (ilv 1) cells in the repressing medium were carrying the plasmid at the time of harvest. In the third experiment a strain, D160-2C, of S. cerevisiae having an arg 6 nonsense mutation was transformed with the i1v 1 plasmid. It was found that 80% more transformed than untransforrned colonies grew on media without Arg, indicating the presence of the gene on the i1v I plasmid.
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