Analysis of a method for selecting temperature-sensitive mutants of BHK cells (original) (raw)

Abstract

A procedure for the isolation of temperature sensitive mutants of BHK cells is described. Mutagenized cells were synchronized in G1 by serum starvation at 33°, and, following release, shifted to 37.5° in the presence of FUdR, to kill cells entering the DNA synthetic phase. Eight independently mutagenized series of cells were carried through four cycles of this selection, and surviving cells were tested for ability to grow at 33° and 39.5° after each cycle. The results suggest that such procedure is effective in enriching for non-leaky ts mutants and favors the isolation of mutants which at 39.5° are inhibited in the processes necessary for DNA synthesis and lose viability rapidly. The effectiveness of repeated selection cycles and the general characteristics of the_ts_ mutants isolated by this method were also evaluated.

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1. Takeharu Nishimoto
   Present address: Cancer Research Institute, Faculty of Medicine, Kyushu University, 812, Fukuoka, Japan

Authors and Affiliations

1. Department of Pathology, New York School of Medicine, 550 First Avenue, 10016, New York, NY
   Takeharu Nishimoto & Claudio Basilico

Authors

1. Takeharu Nishimoto
2. Claudio Basilico

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Nishimoto, T., Basilico, C. Analysis of a method for selecting temperature-sensitive mutants of BHK cells.Somat Cell Mol Genet 4, 323–339 (1978). https://doi.org/10.1007/BF01542846

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• Received: 28 October 1977
• Revised: 19 December 1977
• Issue date: May 1978
• DOI: https://doi.org/10.1007/BF01542846

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