Efficiency of peptide nucleic acid-directed PCR clamping and its application in the investigation of natural diets of the Japanese Eel Leptocephali

Takeshi Terahara, Seinen Chow, Hiroaki Kurogi, Sun Hee Lee, Katsumi Tsukamoto, Noritaka Mochioka, Hideki Tanaka, Haruko Takeyama

研究成果: Contribution to journalArticle査読

22 被引用数 (Scopus)

抄録

Polymerase chain reaction (PCR)-clamping using blocking primer and DNA-analogs, such as peptide nucleotide acid (PNA), may be used to selectively amplify target DNA for molecular diet analysis. We investigated PCR-clamping efficiency by studying PNA position and mismatch with complementary DNA by designing PNAs at five different positions on the nuclear rDNA internal transcribed spacer 1 of the Japanese eel Anguilla japonica in association with intra-specific nucleotide substitutions. All five PNAs were observed to efficiently inhibit amplification of a fully complementary DNA template. One mismatch between PNA and template DNA inhibited amplification of the template DNA, while two or more mismatches did not. DNA samples extracted from dorsal muscle and intestine of eight wild-caught leptochephalus larvae were subjected to this analysis, followed by cloning, nucleotide sequence analysis, and database homology search. Among 12 sequence types obtained from the intestine sample, six were identified as fungi. No sequence similarities were found in the database for the remaining six types, which were not related to one another. These results, in conjunction with our laboratory observations on larval feeding, suggest that eel leptocephali may not be dependent upon living plankton for their food source.

本文言語英語
論文番号e25715
ジャーナルPloS one
6
10
DOI
出版ステータス出版済み - 11 1 2011

All Science Journal Classification (ASJC) codes

  • 一般

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