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In vitro studies on the role of phage T7 gene 4 product in DNA replication

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Summary

A soluble enzyme fraction prepared from T7-infected E. coli is able to initiate DNA synthesis on circular single-stranded phage DNA. The product synthesized in vitro is a full-length linear complementary strand as judged by alkaline sucrose gradient analysis. DNA synthesis requires the products of the phage genes 4 and 5, Mg++, dNTPs and rNTPs; however, ATP by itself can almost completely satisfy the rNTP requirement. The gene 4 product is essential for DNA chain initiation on unprimed single-stranded DNA, but is dispensable for the replication of a ϕX174 DNA-RNA hybrid. The enzyme system from T7-infected cells does not discriminate between the DNA templates from phages ϕX174, M13 or fd and is also capable of replicating native T7 DNA. However, a striking difference with regard to the template DNA is revealed by complementation analysis. Extracts of T7 mutant-infected cells complement each other only with T7 DNA but not with ϕX174 DNA as template.

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Abbreviations

rNTP:

ribonucleoside triphosphate

dNTP:

deoxyribonucleoside triphosphate

BSA:

bovine serum albumin

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Authors and Affiliations

  1. Abteilung Schuster, Max-Planck-Institut für Molekulare Genetik, Berlin-Dahlem, Germany

    Eberhard Scherzinger & Frank Litfin

Authors
  1. Eberhard Scherzinger
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  2. Frank Litfin
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Communicated by W. Arber

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Scherzinger, E., Litfin, F. In vitro studies on the role of phage T7 gene 4 product in DNA replication. Molec. Gen. Genet. 135, 73–86 (1974). https://doi.org/10.1007/BF00433903

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