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基于线粒体16S rRNA基因部分序列等位基因特异性聚合酶链反应快速鉴定北方毛虾(中国毛虾)的起源

Rapid Origin Determination of the Northern Mauxia Shrimp (Acetes chinensis) Based on Allele Specific Polymerase Chain Reaction of Partial Mitochondrial 16S rRNA Gene.

作者信息

Kang Jung-Ha, Noh Eun-Soo, Park Jung-Youn, An Chel-Min, Choi Jung-Hwa, Kim Jin-Koo

机构信息

Fisheries Resources Research Division, NFRDI, Busan 619-705, Korea .

Department of Marine Biology, Pukyung National University, Busan 608-737, Korea .

出版信息

Asian-Australas J Anim Sci. 2015 Apr;28(4):568-72. doi: 10.5713/ajas.14.0613.

DOI:10.5713/ajas.14.0613
PMID:25656197
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4341107/
Abstract

Acetes chinensis is an economically important shrimp that belongs to the Sergestidae family; following fermentation, A. chinensis' economic value, however, is low in China, and much of the catch in China is exported to Korea at a low price, thus leading to potential false labeling. For this reason, we developed a simple method to identify A. chinensis' origin using allele-specific polymerase chain reaction (PCR). Ten single nucleotide polymorphisms (SNPs) were identified from partial (i.e., 570 bp) DNA sequence analysis of the mitochondrial 16s rRNA gene in 96 Korean and 96 Chinese individual shrimp. Among 10 SNP sites, four sites were observed in populations from both countries, and two sites located in the middle with SNP sites at their 3'-ends were used to design allele-specific primers. Among the eight internal primers, the C220F primer specific to the Chinese A. chinensis population amplified a DNA fragment of 364 bp only from that population. We were able to identify the A. chinensis population origin with 100% accuracy using multiplex PCR performed with two external primers and C220F primers. These results show that the 16S rRNA gene that is generally used for the identification of species can be used for the identification of the origin within species of A. chinensis, which is an important finding for the fair trade of the species between Korea and China.

摘要

中国毛虾是一种具有重要经济价值的虾类,属于 Sergestidae 科;然而,在中国,经过发酵后,中国毛虾的经济价值较低,在中国捕获的大部分毛虾都以低价出口到韩国,从而导致潜在的标签错误。因此,我们开发了一种使用等位基因特异性聚合酶链反应(PCR)来鉴定中国毛虾产地的简单方法。通过对 96 只韩国毛虾和 96 只中国毛虾的线粒体 16s rRNA 基因的部分(即 570 bp)DNA 序列分析,鉴定出了 10 个单核苷酸多态性(SNP)。在这 10 个 SNP 位点中,在两国的种群中都观察到了 4 个位点,并且使用位于中间且 3' 端有 SNP 位点的 2 个位点来设计等位基因特异性引物。在这 8 个内部引物中,中国毛虾种群特有的 C220F 引物仅从该种群中扩增出了 364 bp 的 DNA 片段。使用两个外部引物和 C220F 引物进行多重 PCR,我们能够以 100% 的准确率鉴定中国毛虾的种群来源。这些结果表明,通常用于物种鉴定的 16S rRNA 基因可用于中国毛虾种内产地的鉴定,这对于该物种在韩国和中国之间的公平贸易是一项重要发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/b4e6a93779c0/ajas-28-4-568f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/b4f548104408/ajas-28-4-568f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/a70d2748faea/ajas-28-4-568f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/b4e6a93779c0/ajas-28-4-568f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/b4f548104408/ajas-28-4-568f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/a70d2748faea/ajas-28-4-568f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc03/4341107/b4e6a93779c0/ajas-28-4-568f3.jpg

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