(Anaerobic metabolism of aromatic compounds by phototrophic bacteria: biochemical aspects): Annual progress report, April 1988--March 1989

Intensive efforts to define a protocol for generating transposon mutations in R. palustris have continued. An indirect approach, in which a transposon is introduced into a pLAFR1 cosmid containing an approximately 20 kb fragment of R. palustris DNA from a lambda derivative containing Tn5, appears promising. Both plasmids can be introduced into E. coli, and the antibiotic resistances coded for by each subsequently mated into wild-type R. palustris. A mutant isolated following chemical mutagenesis which is unable to grow aerobically on 4-OH benzoate (CGA033) has been complemented biochemically by the same clone which functions in the absence of the transposon; in addition, and more importantly, this indirect tranposon mutagenesis appears to yield mutants with novel phenotypes affecting the anaerobic pathway for 4-OH benzoate utilization. 2 refs.