Design and testing of a functional group-specific DNA probe for the study of natural populations of acetogenic bacteria.
The acetogens, although phylogenetically diverse, can be characterized by their possession of the acetyl coenzyme A (acetyl-CoA) pathway for autotrophic CO2 fixation. The gene encoding formyltetrahydrofolate synthetase, a key enzyme of the acetyl-CoA pathway, was previously cloned from the thermophilic acetogen Clostridium thermoaceticum and has now been tested as a group-specific probe for acetogens. Stable hybrids were formed between the probe and single DNA fragments from eight known acetogens representing six genera. A hybrid was also formed between the probe and a DNA fragment from one sulfate reducer known to be capable of both autotrophic CO2 fixation and acetate catabolism. No such hybrid was formed between the probe and DNA from a homoacetate fermenter not known to use the acetyl-CoA pathway, with two known formyltetrahydrofolate synthetase-producing purine fermenters, or with DNA from 27 other species representing 16 genera of organisms that do not use the acetyl-CoA pathway. DNA purified from cells extracted from horse manure was also screened with the acetogen probe. Six hybrids, indicating at least six detectable acetogen "strains," were observed.
Lovell
C R
CR
Department of Biological Sciences, University of South Carolina, Columbia 29208.
Hui
Y
Y
D000105:Acetyl Coenzyme A / Q000235:genetics*; D001419:Bacteria / Q000235:genetics*; D015342:DNA Probes / Q000138:chemical synthesis; D004269:DNA, Bacterial / Q000302:isolation & purification; D005574:Formate-Tetrahydrofolate Ligase / Q000235:genetics; D005818:Genetic Engineering; D008372:Manure; D009693:Nucleic Acid Hybridization
D015342:DNA Probes; D004269:DNA, Bacterial; D008372:Manure; D000105:Acetyl Coenzyme A; D005574:Formate-Tetrahydrofolate Ligase