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为了对日本进境的冻鲭鱼体内分离出的异尖科线虫进行研究,试验采用寄生虫通用引物NC5和NC2 PCR扩增其核糖体DNA(rDNA)的内转录间隔区(ITS)序列,并进行克隆、转化、测序和序列分析。结果表明:扩增的异尖科线虫样品的ITS序列片段大小为900 bp,包含部分的18S、28S及全部的ITS1(373 bp)、5.8S(159 bp)和ITS2(272 bp)序列,ITS1和ITS2序列与GenBank中抹香鲸异尖线虫(Anisakis physeteris)同源性均为100%,与其他线虫的相似性较低;从鲭鱼中分离到的线虫ITS1和ITS2均与抹香鲸异尖线虫处于同一分支。
In order to study the heteropterans isolated from Japanese frozen mackerel, we sequenced the internal transcribed spacer (ITS) sequences of ribosomal DNA (rDNA) by using the universal primers NC5 and NC2 Cloning, transformation, sequencing and sequence analysis. The results showed that the amplified fragment of ITS sequence was 900 bp in length, including 18S, 28S, ITS1 (373 bp), 5.8S (159 bp) and ITS2 (272 bp) And ITS2 sequences share 100% homology with Anisakis physeteris in GenBank, and have similarities with other nematodes. The ITS1 and ITS2 isolates from mackerel are all in the same Branch.