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

AUTOR(ES)

Shizuya, Hiroaki

RESUMO

We have developed a cell-free system to study bacteriophage λ DNA replication. Maximal DNA synthesis in vitro requires the four deoxynucleoside triphosphates, ATP, and exogenous λ DNA. DNA synthesis requires the products of the phage O and P genes but is not inhibited by λ 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 λO+P+-infected cell extracts will complement extracts prepared from λO- or λP--infected cells.

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