A method for constructing single-copy lac fusions in Salmonella typhimurium and its application to the hemA-prfA operon.

摘要: This report describes a set of Escherichia coli and Salmonella typhimurium strains that permits the reversible transfer lac fusions between plasmid either bacterial chromosome. The system relies on homologous recombination in an E. recD host for from to strain carries S. put operon inserted into trp, resulting are form trp::put::[Kanr-X-lac], where X is promoter or gene fragment under study. homology flanks fusion segment, so can be transduced typhimurium, replacing resident operon. Subsequent transduction with large chromosomal deletion covering allows selection recombinants inherit plasmid. A transposable version was constructed used direct novel locations, including F ara locus. Transductional crosses bearing different segments hemA-prfA were determine contribution hemA region expression prfA other genes downstream typhimurium.