Abstract
Extensive replication of duplex T7 DNA is catalyzed in reactions contining T7 DNA polymerase, T7 gene 4 protein, and T7 RNA polymerase. When the product of this reaction is analyzed in the electron microscope, many eye form and Y form replication intermediates are observed. Replication in vitro is not initiated at a single region of the T7 genome. However, we tentatively conclude that initiation does occur preferentially at a few specific sites along the DNA, and that these sites may be near promoters at which the T7 RNA polymerase initiates transcription.
Highlights
Extensive replication of duplex T7 DNA is catalyzed (Accurate Chemical Corp.) +X174 viral and R F DNA were gifts from in reactions containing T7 DNA polymerase, T7 gene 4 Dr Robert Bambara (Universityof Rochester)
We tentatively conclude that initiation does occur preferentially at a few specific sites along the DNA, and that these sites may be near promoters at which the T7 RNA polymerase initiates transcription
In the preceding paper [5] we describe an in vitro system of each),6 pI of cytochrome c
Summary
In this paperwe present an electronmicro- had been coated with collodion within the previous 24 h. The grids graphic analysisof the product of this in vitro reaction. EXPERIMENTALPROCEDURES photographed with an AEI EM801or Philips 200 electron microscope. I f not otherwise indicated, materials and methods were as specified in the accompanying paper [5]. A t magnifications of 4,000 to lO,oOO, enlarged and measured with an electronic planimeter (Numonics Graphics Calculator,model 250117). This paper has been assigned Report UR-3490-1786. The costs of puhlication of this article were defrayed in part by the payment of page charges. This article must be hereby marked "nduertisement" in accordance with 18 U.S.C. Section 1734 so1el.y to indicate this fact
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