Construction and characterization of an Escherichia coli plasmid bearing a functional gene G of bacteriophage φX174

M. Z. Humayun, R. W. Chambers

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Abstract

In order to study the mutagenic effects of site-specific, covalent modifications of biologically active DNA, we need host cells that are permissive for any type of mutation that might be produced in vivo from the modified DNA. Specifically, we require a general, in vivo complementation system for the bacteriophage ΦX174 gene G, an essential gene that we have chosen for our initial studies of chemical mutagenesis. Toward this end, we have constructed a plasmid (pΦXG) that carries a functional copy of ΦX174 gene G. Three different bacterial strains that are nonpermissive for am9, a gene G amber mutant, have been transformed with pΦXG. The transformants are now permissive for this gene G mutant, but not for the gene A or E mutants that have been tested. This paper describes the construction and the biochemical characterization of this plasmid, pΦXG, and describes some of the biological properties exhibited by the pΦXG-bearing strains.

Original languageEnglish (US)
Pages (from-to)774-778
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume75
Issue number2
DOIs
StatePublished - 1978
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • General

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