基于PCR-RFLP的鼠鹿线粒体12S rRNA基因序列鉴定

Molecular biology international Pub Date : 2013-01-01 Epub Date: 2013-12-23 DOI:10.1155/2013/783925
Chandra Mohan Siddappa, Mohini Saini, Asit Das, Ramesh Doreswamy, Anil K Sharma, Praveen K Gupta
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引用次数: 10

摘要

线粒体12S rRNA已被证明是一种有用的分子标记,可以更好地保护和管理濒危物种。线粒体12S rRNA基因的聚合酶链反应-限制性片段长度多态性(PCR-RFLP)已被证明是鉴定鹿科不同印度鹿种的一种可靠和有效的工具。本研究对家兔科鼠鹿(Moschiola indica)线粒体12S rRNA基因序列进行了计算机分析和表征,以供物种鉴定之用。从毛囊中分离基因组DNA,用通用引物扩增线粒体12S rRNA基因。PCR产物首次克隆并测序。鼠鹿与梅花鹿、桑巴鹿、猪鹿、麝鹿、赤鹿和狗尾鹿的同源性分别为90.04、90.08、90.04、91.2、90.04和90.08%。Lasergene (DNAstar Inc., Madison, WI, USA)的限制性内切图谱显示,使用RsaI、DdeI、BsrI和BstSFI,小鼠鹿线粒体12S rRNA基因序列可以在PCR-RFLP中与其他鹿物种区分。利用预测的模式,可以利用多种生物材料的基因组DNA对鼠鹿进行鉴定,从而为野生动物法医鉴定和物种保护提供分子辅助。
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Sequence Characterization of Mitochondrial 12S rRNA Gene in Mouse Deer (Moschiola indica) for PCR-RFLP Based Species Identification.

Mitochondrial 12S rRNA has proven to be a useful molecular marker for better conservation and management of the endangered species. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the mitochondrial 12S rRNA gene has proven to be a reliable and efficient tool for the identification of different Indian deer species of family cervidae. In the present study, mitochondrial 12S rRNA gene sequence of mouse deer (Moschiola indica) belonging to the family Tragulidae was characterized and analysed in silico for its use in species identification. Genomic DNA was isolated from the hair follicles and mitochondrial 12S rRNA gene was amplified using universal primers. PCR product was cloned and sequenced for the first time. The sequence of mouse deer showed 90.04, 90.08, 90.04, 91.2, 90.04, and 90.08% identities with sika deer, sambar, hog deer, musk deer, chital, and barking deer, respectively. Restriction mapping in Lasergene (DNAstar Inc., Madison, WI, USA) revealed that mouse deer mitochondrial 12S rRNA gene sequence can be differentiated from the other deer species in PCR-RFLP using RsaI, DdeI, BsrI, and BstSFI. With the help of predicted pattern, mouse deer can be identified using genomic DNA from a variety of biomaterials, thereby providing molecular aid in wildlife forensics and conservation of the species.

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