Transposon Mutagenesis of Rhodobacter sphaeroides

 

Timothy Paustian and Robin Kurtz

Tested Studies in Laboratory Teaching, 1994, Volume 15

Abstract

Creation and characterization of mutants is the basis for any genetic analysis. This exercise demonstrates a simple, safe procedure for transposon mutagenesis of Rhodobacter sphaeroides, a purple non-sulfur photosynthetic bacteria. Students perform the mutagenesis by mating a transposon-carrying plasmid from Escherichia coli to R. sphaeroides and then selecting for the drug resistance carried on the transposon. Only R. sphaeroides carrying the transposon in the chromosome survive the selection. Transposon carrying mutants are then scored for various phenotypes. The metabolic diversity of R. sphaeroides allows the isolation of nutritional, photopigment, and photosynthetic mutants. Further analysis of mutants is possible.

Keywords:  bacteria, mutagenesis, conjugation, transposon

University of Toronto (1993)