Rapid differentiation of hyperthermophilic Archaea by restriction mapping of the intergenic spacer regions of the ribosomal RNA operons.
We report a sensitive and convenient method for rapid differentiation of new isolates of hyperthermophilic Archaea. Polymerase chain reaction (PCR) was used to amplify the intergenic spacer regions of the ribosomal RNA operons of eight Archaea. Spacer regions from one Euryarcheote, Pyrococcus furiosus, and one Crenarcheote, Pyrodictium brockii, were sequenced completely. Restriction fragment length polymorphism (RFLP) analyses were performed on the spacer regions from eight hyperthermophilic Archaea, and the restriction patterns were used as fingerprints for six known strains and two isolates. The PCR-RFLP method used in this study allowed the differentiation of seven of the eight strains tested and could be generally applicable to all the Archaea.
DiRuggiero
J
J
Center of Marine Biotechnology, University of Maryland System, Baltimore 21202, USA.
Tuttle
J H
JH
Robb
F T
FT
D001105:Archaea / Q000145:classification*; D001483:Base Sequence; D004275:DNA, Ribosomal / Q000235:genetics*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012335:RNA, Ribosomal / Q000235:genetics*; D015183:Restriction Mapping; D017421:Sequence Analysis; D013997:Time Factors
D004275:DNA, Ribosomal; D012335:RNA, Ribosomal