Induction and isolation of DNA transformation mutants in the yeast Saccharomyces cerevisiae

Conference · Thu Jan 01 04:00:00 EST 1987 · Proc. Soc. Exp. Biol. Med.; (United States)

OSTI ID:5517670

Hegerich, P A; Bruschi, C V

The objective of this research was to induce and isolate mutants of the yeast Saccharomyces cerevisiae which have become transformable by purified plasmid DNA. Non-transformable yeast cells were mutagenized by ultraviolet light using a 65% lethal dose (480 ergs/mm/sup 2/). After a period of overnight liquid holding recovery, the irradiated cells were subjected to DNA transformation using our CaCl/sub 2/ protocol with the multi-marker shuttle plasmid pBB carrying the LEU 2 leucine gene. Following transformation the colonies that grew on selective leucineless medium were identified and subjected to further genetic analysis. From a total of 1 x 10/sup 9/ cells the authors have isolated 7 colonies deriving from putative mutants that have acquired the capability to uptake plasmid DNA. The transformants were cured from the plasmid by its mitotic loss on non-selective medium, then re-transformed to verify their genetic competence to give rise to a number of transformants comparable to transformable strains. We have identified and isolated one mutant, coded trs-1, which is able to reproduce a frequency of transformation comparable with the tranformable control. They, therefore, conclude that this mutant is specific for plasmid DNA transformation and that the mutation is mitotically stable.

• Research Organization: East Carolina Univ. School of Medicine, Greenville, NC
• OSTI ID: 5517670
• Report Number(s): CONF-8611204-
• Conference Information: Journal Name: Proc. Soc. Exp. Biol. Med.; (United States) Journal Volume: 184:4
• Country of Publication: United States
• Language: English

Related Subjects

560130* -- Radiation Effects on Microorganisms
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACIDS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
DNA
ELECTROMAGNETIC RADIATION
FUNGI
GENES
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
IRRADIATION
LETHAL IRRADIATION
LEUCINE
MICROORGANISMS
MUTANTS
MUTATION FREQUENCY
MUTATIONS
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
PLASMIDS
RADIATION EFFECTS
RADIATIONS
RADIOINDUCTION
SACCHAROMYCES
SACCHAROMYCES CEREVISIAE
ULTRAVIOLET RADIATION
YEASTS