Investigation on Natural Diets of Larval Marine Animals Using Peptide Nucleic Acid-Directed Polymerase Chain Reaction Clamping

Seinen Chow, Sayaka Suzuki, Tadashi Matsunaga, Shane Lavery, Andrew Jeffs, Haruko Takeyama

    Research output: Contribution to journalArticle

    36 Citations (Scopus)

    Abstract

    The stomach contents of the larvae of marine animals are usually very small in quantity and amorphous, especially in invertebrates, making morphological methods of identification very difficult. Nucleotide sequence analysis using polymerase chain reaction (PCR) is a likely approach, but the large quantity of larval (host) DNA present may mask subtle signals from the prey genome. We have adopted peptide nucleic acid (PNA)-directed PCR clamping to selectively inhibit amplification of host DNA for this purpose. The Japanese spiny lobster (Panulirus japonicus) and eel (Anguilla japonica) were used as model host and prey organisms, respectively. A lobster-specific PNA oligomer (20 bases) was designed to anneal to the sequence at the junction of the 18 S rDNA gene and the internal transcribed spacer 1 (ITS1) of the lobster. PCR using eukaryote universal primers for amplifying the ITS1 region used in conjunction with the lobster-specific PNA on a mixed DNA template of lobster and eel demonstrated successful inhibition of lobster ITS1 amplification while allowing efficient amplification of eel ITS1. This method was then applied to wild-caught lobster larvae of P. japonicus and P. longipes bispinosus collected around Ryukyu Archipelago, Japan. ITS1 sequences of a wide variety of animals (Ctenophora, Cnidaria, Crustacea, Teleostei, Mollusca, and Chaetognatha) were detected.

    Original languageEnglish
    Pages (from-to)305-313
    Number of pages9
    JournalMarine Biotechnology
    Volume13
    Issue number2
    DOIs
    Publication statusPublished - 2011 Apr

    Fingerprint

    Peptide Nucleic Acids
    Eels
    nucleic acid
    lobster
    Palinuridae
    lobsters
    Constriction
    peptide
    nucleic acids
    polymerase chain reaction
    Panulirus japonicus
    internal transcribed spacers
    peptides
    diet
    Diet
    Polymerase Chain Reaction
    Larva
    animal
    DNA
    Ctenophora

    Keywords

    • Gut content analysis
    • Larval diet
    • PCR-clamping
    • PNA
    • Spiny lobster

    ASJC Scopus subject areas

    • Aquatic Science
    • Biotechnology

    Cite this

    Investigation on Natural Diets of Larval Marine Animals Using Peptide Nucleic Acid-Directed Polymerase Chain Reaction Clamping. / Chow, Seinen; Suzuki, Sayaka; Matsunaga, Tadashi; Lavery, Shane; Jeffs, Andrew; Takeyama, Haruko.

    In: Marine Biotechnology, Vol. 13, No. 2, 04.2011, p. 305-313.

    Research output: Contribution to journalArticle

    Chow, Seinen ; Suzuki, Sayaka ; Matsunaga, Tadashi ; Lavery, Shane ; Jeffs, Andrew ; Takeyama, Haruko. / Investigation on Natural Diets of Larval Marine Animals Using Peptide Nucleic Acid-Directed Polymerase Chain Reaction Clamping. In: Marine Biotechnology. 2011 ; Vol. 13, No. 2. pp. 305-313.
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