Design and evaluation of novel primers for the detection of genes encoding diverse enzymes of methylotrophy and autotrophy.
The phylogenetic significance of the diversity of key enzymes of methylotrophic and autotrophic metabolism is discussed. Primers for these key enzymes were designed using gene sequences encoding methanol dehydrogenase (mxaF; using subsets from database sequences for 22 Bacteria), hydroxypyruvate reductase (hpr; 36 sequences), methylamine dehydrogenase (mauA; 12 sequences), methanesulfonate monooxygenase (msmA; four sequences), and the ccbL and cbbM genes of ribulose bisphosphate carboxylase (26 and 23 sequences). These were effective in amplifying the correct gene products for the target genes in reference organisms and in test organisms not previously shown to contain the genes, as well as in some methylotrophic Proteobacteria isolated from the human mouth. The availability of the new primers increases the probability of detecting diverse examples of the genes encoding these key enzymes both in natural populations and in isolated bacterial strains.
King's College London Dental Institute, Microbiology, Guy's Campus, London, SE1 9RT, UK.
Wei-Lian
WL
Hung
William G
WG
Wade
Yin
Y
Chen
Donovan P
DP
Kelly
Ann P
AP
Wood
D000429:Alcohol Oxidoreductases / Q000235:genetics; D052818:Autotrophic Processes; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D002244:Carbon / Q000378:metabolism*; D017931:DNA Primers; D014644:Genetic Variation; D006801:Humans; D050537:Hydroxypyruvate Reductase / Q000235:genetics; D009055:Mouth / Q000382:microbiology; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D002244:Carbon; D000429:Alcohol Oxidoreductases; D050537:Hydroxypyruvate Reductase; C019641:alcohol dehydrogenase (acceptor); C021567:methylamine dehydrogenase; D000587:Oxidoreductases Acting on CH-NH Group Donors