| 摘要: |
| 【目的】探索从林麝粪便中提取基因组DNA并进行物种鉴定的可行性，为林麝野外调查工作以及分子粪便学在林麝保护中的应用提供科学依据。【方法】采集林麝粪便，采用改良的CTAB抽提基因组DNA；按常规PCR法对mtDNA D-loop区和Cyt-b基因进行扩增测序后，用DNAMAN 6.0软件进行系统发育分析；在不同温度存放不同时间，研究林麝粪便基因组DNA的提取与mtDNA D-loop区和Cyt-b基因PCR扩增的时效性。【结果】以提取的基因组DNA为模板进行PCR扩增均能特异性地扩增出mtDNA D-loop区和Cyt-b基因。系统发育分析结果表明：所扩增产物为林麝的mtDNA D-loop区和Cyt-b基因。时效性分析结果显示：常温和4℃条件下保存96 h以内的林麝粪便所提取的基因组DNA能有效扩增出mtDNA D-loop区和Cyt-b基因。【结论】自然条件下林麝粪便中林麝自身体细胞DNA保存时间较长，通过在野外采集林麝粪便提取基因组DNA并进行物种鉴定是可行的。 |
| 关键词: 林麝 粪便 线粒体DNA D-loop区 Cyt-b基因 |
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| 基金项目:四川省林业厅省级自然保护区重点物种科学研究项目；四川农业大学院级专项 |
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| Extraction and PCR amplification of genomic DNA of forest musk deer feces |
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KANG Qiang,ZHOU Xin,LUO Yan,et al
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| Abstract: |
| 【Objective】This study explored the feasibility of genomic DNA extraction and species identification from forest musk deer feces to provide scientific basis for field investigation and application of deer molecular scatology in protection of forest musk deer.【Method】Genomic DNA of forest musk deer feces was extracted by the modified CTAB extraction method.Cyt-b gene and mtDNA D-loop region were amplified by PCR.Both sequences were analyzed by DNAMAN 6.0 software.The timeliness of genomic DNA extraction and PCR amplification of mtDNA D-loop region and Cyt-b gene was investigated at different temperatures.【Result】Genomic DNA of forest musk deer feces could be extracted,and the phylogenetic analysis showed that both mtDNA D-loop region and Cyt-b gene could be amplified.Timeliness analysis showed that mtDNA D-loop region and Cyt-b gene could be specifically amplified within 96 h at both room temperature and 4 ℃.【Conclusion】The DNA of musk deer in feces can be preserved at a long time under natural conditions.It is feasible to conduct species identification by collecting forest musk deer feces in the wild. |
| Key words: forest musk deer feces genomic DNA D-loop region Cyt-b gene |
