Synthesis of Bacteriophage Lambda DNA In Vitro: Requirement for O and P Gene Products

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Abstract
We have developed a cell-free system to study bacteriophage lambda DNA replication. Maximal DNA synthesis in vitro requires the four deoxynucleoside triphosphates, ATP, and exogenous lambda DNA. DNA synthesis requires the products of the phage O and P genes but is not inhibited by lambda repressor. The kinetics of synthesis is linear for 10-15 min; however, the product of synthesis amounts to only 0.5-1% of the added template DNA. As judged by isopycnic analysis, extensive regions of the template are copied. Sedimentation analysis indicates that all of the product consists of short (11S) DNA chains. Fractions partially purified from lambdaO(+)P(+)-infected cell extracts will complement extracts prepared from lambdaO(-) or lambdaP(-)-infected cells.