Isolation of yeast DNA replication mutants in permeabilized cells.

摘要: A random population of temperature-sensitive mutants was screened by assaying for defects in DNA synthesis a permeabilized yeast replication system. Twenty defective vitro have been isolated. In this paper we describe eight these mutants. Seven them fall into three complementation groups--cdc2, cdc8, and cdc16--involved the control cell-division cycle. Because represents propagation forks active vivo at time permeabilization, our finding that cdc2 cdc16 can incorporate dTMP such cells 23 degrees C but not 37 supports conclusion two mutations directly affect forks. Such an involvement previously suggested analysis CDC2 less clear CDC16. Finally, usefulness screening procedure is demonstrated isolation undescribed groups. One strain shows serious defect normal RNA synthesis.