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Nucleotide Sequence of the SrRNA Gene of Entamoeba gingivalis : Applications for Construction of a Species‐Specific DNA Probe and Phylogenetic Analysis
Author(s) -
Yamamoto Ayako,
Kikuta Noriko,
Hashimoto Tetsuo,
Oyaizu Hiroshi,
Goto Nobuichi
Publication year - 1995
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1995.tb02187.x
Subject(s) - biology , nucleic acid sequence , genetics , phylogenetic tree , ribosomal rna , gene , entamoeba , microbiology and biotechnology , hybridization probe , oligonucleotide , entamoeba histolytica
The small subunit ribosomal RNA (SrRNA) gene of Entamoeba gingivalis was amplified by PCR and the product of 1.9‐kbp sequence was cloned into a plasmid vector pUC18. Four clones were isolated and sequenced. The insert DNAs were 1918‐ to 1921‐bp long and A+T rich (65.5%). The four SrRNA sequences of E. gingivalis were found to be aligned with those of nine related protozoans while searching for E. gingivalis ‐specific sequences. A sequence of 28 oligonucleotides was chosen, chemically synthesized, and labeled with digoxigenin for use as a DNA probe. The probe thus constructed was shown to hybridize only with either the SrRNA‐coding DNAs or the cells of the two E. gingivalis strains and not with those of other protozoans or oral fungi tested. A representative SrRNA‐sequence was analyzed and a phylogenetic tree was constructed by the neighbor‐joining (NJ) method. Among the protists examined, E. gingivalis was placed next to Entamoeba histolytica as expected from the traditional taxonomy.