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PCR Amplification and Sequence Analysis of ITS rDNA of Anisakid larvae in Imported Pacific Cod

Hong Qing-lin

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Abstract

The aims of this study were to amplify and analyze the sequence of internal transcribed spacers(ITS)of ribosomal DNA(rDNA)of Anisakid larvae samples isolated from frozen pacific cod imported from Japan.The DNA of the samples were extracted and the ITS sequences were amplified by PCR with primers NC5and NC2.The products were cloned into pMD18-T vector and the inserts were successfully sequenced.The results revealed ITS inserts were 906bp in length and consisted of partial 18S,28Sand complete ITS1(353bp),5.8S(157bp)and ITS2(299bp)rDNA sequences.The similarity in ITS1and ITS2sequences among the Anisakid larvae sample and Pseudoterranova decipiens available in GenBank were over 99.7%,and lower homology with other nematodes.The results of the present study provided a foundation for further studies of Anisakid larvae.

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What this paper is about

The aims of this study were to amplify and analyze the sequence of internal transcribed spacers(ITS)of ribosomal DNA(rDNA)of Anisakid larvae samples isolated from frozen pacific cod imported from Japan.The DNA of the samples were extracted and the ITS sequences were amplified by PCR with primers NC5and NC2.The products were cloned into pMD18-T vector and the inserts were successfully sequenced.The results revealed ITS inserts were 906bp in length and consisted of partial 18S,28Sand complete ITS1(353bp),5.8S(157bp)and ITS2(299bp)rDNA sequences.The similarity in ITS1and ITS2sequences among the Anisakid larvae sample and Pseudoterranova decipiens available in GenBank were over 99.7%,and lower homology with other nematodes.The results of the present study provided a foundation for further studies of Anisakid larvae.

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Available abstract

The aims of this study were to amplify and analyze the sequence of internal transcribed spacers(ITS)of ribosomal DNA(rDNA)of Anisakid larvae samples isolated from frozen pacific cod imported from Japan.The DNA of the samples were extracted and the ITS sequences were amplified by PCR with primers NC5and NC2.The products were cloned into pMD18-T vector and the inserts were successfully sequenced.The results revealed ITS inserts were 906bp in length and consisted of partial 18S,28Sand complete ITS1(353bp),5.8S(157bp)and ITS2(299bp)rDNA sequences.The similarity in ITS1and ITS2sequences among the Anisakid larvae sample and Pseudoterranova decipiens available in GenBank were over 99.7%,and lower homology with other nematodes.The results of the present study provided a foundation for further studies of Anisakid larvae.

Key concepts: GenBank, Biology, Ribosomal DNA, Homology (biology), Larva, Polymerase chain reaction, DNA sequencing, Sequence analysis

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PCR Amplification and Sequence Analysis of ITS rDNA of Anisakid larvae in Imported Pacific Cod — Research Paper | ScholarLens