i. Why was the shift in temperature an important step of the experimental protocol?

Human Anatomy & Physiology (11th Edition)
11th Edition
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Author:Elaine N. Marieb, Katja N. Hoehn
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Chapter1: The Human Body: An Orientation
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8. In genetic screens of mutagenized yeast, temperature sensitive mutants defective in the
secretory pathway (sec mutants) were identified. These mutants have helped scientists
study the protein secretion pathway that is necessary/essential for life in eukaryotic
cells.
To identify sec mutants: mutagenized yeast cells were grown for 12 hours at low
temperature, shifted to higher temperature for two hours, and then the population of
cells were centrifuged in a density gradient.
a. Mutagenized cells were first grown at the lower temperature and then shifted to
the higher temperature.
i. Why was the shift in temperature an important step of the experimental
protocol?
ii. Why would the experiment have been unsuccessful if the cells were
grown exclusively at the higher temperature?
b. After centrifugation, where would sec mutant cells be located compared to other
cells in the population? Briefly explain why.
Transcribed Image Text:8. In genetic screens of mutagenized yeast, temperature sensitive mutants defective in the secretory pathway (sec mutants) were identified. These mutants have helped scientists study the protein secretion pathway that is necessary/essential for life in eukaryotic cells. To identify sec mutants: mutagenized yeast cells were grown for 12 hours at low temperature, shifted to higher temperature for two hours, and then the population of cells were centrifuged in a density gradient. a. Mutagenized cells were first grown at the lower temperature and then shifted to the higher temperature. i. Why was the shift in temperature an important step of the experimental protocol? ii. Why would the experiment have been unsuccessful if the cells were grown exclusively at the higher temperature? b. After centrifugation, where would sec mutant cells be located compared to other cells in the population? Briefly explain why.
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