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用于四种蛤类物种鉴定的16S rRNA基因片段的聚合酶链反应-限制性片段长度多态性分析

Polymerase chain reaction-restriction fragment length polymorphism analysis of a 16S rRNA gene fragment for authentication of four clam species.

作者信息

Fernandez Alicia, García Teresa, Gonzalez Isabel, Asensio Luis, Rodriguez Miguel Angel, Hernández Pablo E, Martin Rosario

机构信息

Departamento de Nutrición y Bromatología III, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.

出版信息

J Food Prot. 2002 Apr;65(4):692-5. doi: 10.4315/0362-028x-65.4.692.

DOI:10.4315/0362-028x-65.4.692
PMID:11952222
Abstract

Specific identification of four clam species, Ruditapes decussatus (grooved carpet shell), Venerupis pullastra (pullet carpet shell), Ruditapes philippinarum (Japanese carpet shell), and Venerupis rhomboides (yellow carpet shell), was achieved by polymerase chain reaction-restriction fragment length polymorphism analysis of a fragment of the mitochondrial 16S rRNA gene. Amplification of DNA isolated from the foot muscle produced fragments of 511 bp for V. pullastra, 523 bp for R. decussatus, 545 bp for R. philippinarum, and 502 bp for V. rhomboides. The restriction profiles obtained by agarose gel electrophoresis when amplicons were digested with endonucleases BsmAI and BsrI allowed unequivocal identification of the four clam species. This approach would be less costly, simpler, and quicker than conventional sequencing of polymerase chain reaction products followed by detailed comparison of individual sequences, especially when large numbers of samples need to be analyzed.

摘要

通过对线粒体16S rRNA基因片段进行聚合酶链反应-限制性片段长度多态性分析,实现了对四种蛤蜊物种的特异性鉴定,这四种蛤蜊分别是:波纹巴非蛤(Ruditapes decussatus,有沟波纹蛤)、紫彩血蛤(Venerupis pullastra,小型血蛤)、菲律宾蛤仔(Ruditapes philippinarum,菲律宾蛤)和菱形紫蛤(Venerupis rhomboides,黄海紫蛤)。从足部肌肉中分离的DNA经扩增后,紫彩血蛤产生511 bp的片段,波纹巴非蛤产生523 bp的片段,菲律宾蛤仔产生545 bp的片段,菱形紫蛤产生502 bp的片段。当用内切酶BsmAI和BsrI消化扩增产物后,通过琼脂糖凝胶电泳获得的限制性图谱能够明确鉴定这四种蛤蜊物种。与传统的对聚合酶链反应产物进行测序并随后详细比较各个序列的方法相比,这种方法成本更低、更简单、更快速,尤其是在需要分析大量样本时。

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