Induction and isolation of DNA transformation mutants in the yeast Saccharomyces cerevisiae
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-; TRN: 88-006165
- Journal Information:
- Proc. Soc. Exp. Biol. Med.; (United States), Vol. 184:4; Conference: Annual meeting of the Southeast Section of the Society for Experimental Biology and Medicine, Chapel Hill, NC, USA, 13 Nov 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
MUTATIONS
RADIOINDUCTION
SACCHAROMYCES CEREVISIAE
GENETIC RADIATION EFFECTS
DNA
GENES
LETHAL IRRADIATION
LEUCINE
MUTANTS
MUTATION FREQUENCY
PLASMIDS
ULTRAVIOLET RADIATION
AMINO ACIDS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
ELECTROMAGNETIC RADIATION
FUNGI
GENETIC EFFECTS
IRRADIATION
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
RADIATION EFFECTS
RADIATIONS
SACCHAROMYCES
YEASTS
560130* - Radiation Effects on Microorganisms