Bacteriophage T7 DNA Synthesis in Isolated DNA-Membrane Complexes
Melvin S. Center
Abstract
Melvin S. Center
Abstract
A DNA-membrane complex isolated from Escherichia coli infected with bacteriophage T7 contains newly synthesized T7 DNA and the T7 DNA polymerase (gene 5 product). The DNA present in the complex appears to exist as a concatemer which contains single-strand breaks and possibly internal single-stranded regions (gaps). The complex is capable of synthesizing T7 DNA by using endogenous template, and part of the DNA is made by a semiconservative mechanism. A portion of the in vitro synthesized DNA sediments in alkaline sucrose as 10-11S material. This DNA is converted to a larger-molecular-weight material after treatment with T4 polynucleotide ligase and E. coli DNA polymerase I.
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A DNA-membrane complex isolated from Escherichia coli infected with bacteriophage T7 contains newly synthesized T7 DNA and the T7 DNA polymerase (gene 5 product). The DNA present in the complex appears to exist as a concatemer which contains single-strand breaks and possibly internal single-stranded regions (gaps). The complex is capable of synthesizing T7 DNA by using endogenous template, and part of the DNA is made by a semiconservative mechanism. A portion of the in vitro synthesized DNA sediments in alkaline sucrose as 10-11S material. This DNA is converted to a larger-molecular-weight material after treatment with T4 polynucleotide ligase and E. coli DNA polymerase I.
Key concepts: DNA clamp, Biology, DNA polymerase II, DNA polymerase, DNA polymerase I, DNA ligase, In vitro recombination, DNA