基于细胞色素氧化酶亚基I（COI）基因和性别决定基因（SRY）基因鉴定梅花鹿、马鹿及其杂交样本

[Identification of Cervus nippon, C.elaphus and their hybridize samples based on COI and SRY gene].

作者信息

Wei Yi-Cong, Jiang Chao, Yuan Yuan, Zhao Yu-Yang, Jin Yan, Huang Lu-Qi

机构信息

College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China.

State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2017 Dec;42(23):4588-4592. doi: 10.19540/j.cnki.cjcmm.20171030.020.

DOI:10.19540/j.cnki.cjcmm.20171030.020

PMID:29376256

Abstract

For rapid identification of Cervus nippon, C. elaphus and their hybridize samples, the specific PCR for mutual authentication of them was established based on the SNPs in COI and SRY sequence. C. nippon, C. elaphus and their hybridize samples were collected from different origins, total DNA of 24 identified samples were extracted, and the COI and SRY gene was seqenced. SNPs in the COI and SRY sequences of the samples were found by Clustul X 2.1 program. Primers for identifying C. nippon and C. elaphus were designed according to the SNP site, two multi-PCR reaction system were established to identify them. In addition, 24 samples which were randomly collected in different herbal medicine market were identified. The band special for C. nippon (232 bp)and band special for C. elaphus (518 bp) based on COI sequence,and the band special for C. nippon (803 bp)and band special for C. elaphus (425 bp) based on SRY sequence, were found using multi-PCR reaction, and three of the twenty-four samples were identified as the hybridize samples. The multi-PCR reaction system could be used to identify C. nippon, C. elaphus and their hybridize samples.

摘要

为快速鉴定梅花鹿、马鹿及其杂交样本，基于细胞色素氧化酶亚基I（COI）和Y染色体性别决定区（SRY）序列中的单核苷酸多态性（SNP），建立了三者相互鉴别的特异性聚合酶链反应（PCR）。从不同来源采集梅花鹿、马鹿及其杂交样本，提取24份已鉴定样本的总DNA，并对COI和SRY基因进行测序。利用Clustul X 2.1程序查找样本COI和SRY序列中的SNP。根据SNP位点设计鉴定梅花鹿和马鹿的引物，建立两个多重PCR反应体系对其进行鉴定。此外，对在不同中药材市场随机采集的24份样本进行鉴定。通过多重PCR反应，发现基于COI序列的梅花鹿特异性条带（232 bp）和马鹿特异性条带（518 bp），以及基于SRY序列的梅花鹿特异性条带（803 bp）和马鹿特异性条带（425 bp），24份样本中有3份被鉴定为杂交样本。该多重PCR反应体系可用于鉴定梅花鹿、马鹿及其杂交样本。