论文总字数：21676字

摘 要

水产品物种掺假在全球导致了经济、食品安全等问题，因此建立一种快速高效的水产品物种鉴定技术迫在眉睫。微型DNA条形码技术可对深加工水产品制品进行物种鉴定，较于普通的DNA条形码技术具有更高鉴定成功率，在水产品物种掺假检测中应用广泛。文中针对国内最具有代表性的33种不同的鱼，基于通用引物实现扩增、测序以及序列比对。

结果发现，选择天根试剂盒在样品中提取DNA操作简单快速，DNA纯度高，且所有样品均可以扩增出约165bp的COI条形码片段，但只有12个品种（35.3%）可在物种水平进行准确鉴定。在序列比对分析时，发现使用DNA条形码（FDB）和微型DNA条形码（MDB）进行物种鉴定的结果无差异，但100bp-300bp左右的短序列更容易扩增出来。

本研究对DNA条形码的目的基因选择以及适宜片段长度进行了研究。为建立适用于我国常见鱼类品种的DNA条形码技术提供了理论依据。本课题的实施可为政府监管部门准确鉴定市场鱼类品种提供准确的途径，起到规范我国水产品市场和保护消费者权益的作用。

关键词：FDB MDB 物种鉴定 COI基因

Comparative study on mini DNA Barcode in Identification of

Common Fish breeds in China

Abstract

Aquatic product species adulteration has led to economic, food safety and other problems in the world, so it is urgent to establish a rapid and efficient aquatic product species identification technology. Micro DNA bar code technology can be used to identify species of deep processed aquatic products, which has higher identification success rate than ordinary DNA bar code technology, and is widely used in species adulteration detection of aquatic products. In the article, for the 33 representative fishes in the country, amplification, sequencing and sequence alignment were performed based on universal primers.

It was found that the selection of Tiangen kit for DNA extraction in samples is simple and rapid, and the DNA purity is high. All samples could amplify about 165bp of COI barcode, but only 12 varieties(35.3%)could be accurately identified at the species level. In sequence comparison analysis, it was found that the results of identification using DNA bar code(FDB)and micro-DNA bar code(MDB)were almost consistent, but the short sequence of about 100bp300bp was more easily amplified.

In the article, the selection of target genes for DNA barcodes and the length of suitable fragments were analyzed. It provides a theoretical basis for the establishment of DNA bar code technology suitable for common fish breeds in China. The implementation of this topic can provide an accurate way for government regulatory authorities to accurately identify market fish species, and play a key role in regulating China's aquatic product market and protecting consumer rights.

Key words: FDB; MDB; Species identification; COI gene

目 录

摘 要 I

Abstract II

第一章 文献综述 1

1.1水产品概述 1

1.2水产品品种掺假 1

1.2.1水产品品种掺假的定义 1

1.2.2鱼类产品品种掺假的现状 1

1.3 鱼类产品品种鉴定技术 2

1.3.1 PCR-RFLP 2

1.3.2 荧光PCR 2

1.3.3 DNA条形码 3

1.3.4微型DNA条形码 4

1.4目的意义 4

第二章 材料与方法 5

2.1样品的采集 5

2.2 主要仪器 6

2.3 主要试剂 6

2.4 实验方法 6

2.4.1 DNA提取 6

2.4.2 DNA浓度和纯度测定 7

2.4.3 总DNA电泳 7

2.4.4 PCR扩增及测序 7

2.4.5 序列比对与分析 8

第三章 实验结果与讨论 9

3.1 DNA提取 9

3.2 PCR扩增与测序 9

3.3 序列比对与分析 11

第四章 结论与展望 16

4.1结论 16

4.2展望 16

参考文献 17

致 谢 19

文献综述

1.1水产品概述

从概念上看，水产品并非单一的产品，而更多的是不同产品的统称，包括通过江河湖海捕捞、养殖等获得的水生动物、植物和相关加工产品。通过归纳总结，我们将水产品划分为两种不同的类型：首先，鲜活水产品，例如养殖或捕捞的虾、鱼、贝、水藻等；其次，加工海产品，例如通过腌制、熏制、冷冻、罐装等处理的产品。

众所周知，水产品对于人类而言是至关重要的，在满足味觉体验的同时，而且营养丰富均衡。相比于其他肉类食品，水产品拥有质量更优、含量更高的蛋白质，深受广大消费者的好评。

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