Phylogenetic Analysis of Marine Picoplankton Using Tau RNA Sequences.
Final progress rept. Jul 90-Feb 91, Annual rept. Feb-Sep 91,
INDIANA UNIV AT BLOOMINGTON
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Phylogenetic identification of picoplankton. Using tangential flow filtration, we collected bulk amounts of marine picoplankton from the central Atlantic Ocean Sargasso Sea, Hydrostation S and northern Pacific Ocean Aloha Station. DNA prepared from both populations was analyzed by hybridization using kingdom-specific probes complementary to 16S rRNA, and determined to be largely eubacteria little eukaryote-specific and little or no archaebacterial-specific DNAs were detected. DNA from the Aloha Station was suitable for cloning experiments and a random library of 20 kb Sau3a fragments in phage gamma EMBL3 was established. This library is available to other investigators upon request. During these past two years, screening that library for 16S rRNA-containing clones, sequencing the genes and carrying out phylogenetic analysis occupied substantial effort. Approximately 50 rRNA gene-containing clones were identified by hybridization and the unique types determined by sequencing using single dideoxynucleotide reactions. Several hundred nucleotides of sequence was determined for each of the unique types.