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Applications of Nucleic Acid Hybridization in Microbial Ecology
Author(s) -
Holben William E.,
Tiedje James M.
Publication year - 1988
Publication title -
ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.144
H-Index - 294
eISSN - 1939-9170
pISSN - 0012-9658
DOI - 10.2307/1941005
Subject(s) - nucleic acid , biology , dna , gene , nucleic acid thermodynamics , organism , computational biology , population , genetics , dna sequencing , rna , microbial ecology , ecology , bacteria , demography , sociology
Nucleic acid hybridization techniques offer a new approach to answer old, intractable questions in microbial ecology as well as new questions. These include characterization of the predominant, yet unculturable populations in nature, the role of the environment in gene expression, and the extent of gene exchange among communities in nature. The essence of this methodology is the denaturing and annealing of complementary strands of nucleic acid molecules. The specificity of this hybridization reaction can be controlled such that only identical, or nearly identical, sequences in a complex mixture of nucleic acids extracted from a population or community can anneal. Labeled DNA or RNA sequences (probes) are introduced into hybridization reactions to identify and quantitate a particular organism containing the complementary target sequence. Methods for the recovery and purification of DNA from soils and sediments are given, as well as important considerations for the selection of probe and target sequences, and for the methods of detection and quantitation. Some advantages of this methodology include the abilities to: (1) detect populations without prior culturing of organisms, (2) detect specific organisms without the need for selectable markers, (3) detect multiple populations in the same analysis, and (4) detect genetic rearrangements or gene transfer in natural communities.