Characterization of Intragenic Recombination in a Hyperthermophilic Archaeon via Conjugational DNA Exchange

Abstract

Sulfolobus acidocaldarius is so far the only hyperthermophilic archaeon in which genetic recombination can be assayed by conjugation and simple selections. Crosses among spontanteous pyr mutants were able to resolve closely spaced chromosomal mutations, identify deletions and rearrangements, and map mutations to a given deletion interval. Frameshift mutations in pyrE exerted polar effects that depressed orotidine-5′-monophosphate decarboxylase activity (encoded by pyrF ), whereas base pair substitutions and an 18-bp deletion had no effect.