| 摘要: |
| 本研究从实验室保藏的节旋藻(Arthrospira)藻种出发, 挑取形态不同的单藻丝体进行纯化培养,采用6 种方法进行全基因组DNA 提取的比较研究, 而后以16S rRNA-ITS 区基因作为分子标记对藻株进行相关序列测定和分子系统进化分析。结果表明, 冻融CTAB 法能够提取出包含染色体外DNA 在内的节旋藻全基因组, 高质量样品可以满足分子生物学实验要求; 分子系统研究表明, 纯化藻株皆为钝顶节旋藻, 节旋藻与螺旋藻在分子鉴定中属间差异明显。 |
| 关键词: 钝顶节旋藻(Arthrospira platensis) 全基因组提取 PCR 16S rRNA-ITS |
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| 基金项目:海洋公益性行业科研专项经费资助项目(200905021-3, 201205027); 山东省自然科学基金项目(ZR2012DQ015); 广东省中国科学院全面战略合作项目(2011A090100040) |
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| Comparative investigation on whole genome DNA extraction of axenic strains of Arthrospira platensis |
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| Abstract: |
| Single filaments of Arthrospira/Spirulina strains were picked up and axenicly cultured. Six protocols for extraction of whole genome DNA of the Arthrospira strains were comparatively studied. The phylogenetic analysis of 16S rRNA-ITS (internally transcribed spacer) gene sequences as molecular markers were conducted. The results show that the method of Freezing-thawing CTAB was effective in extracting extra-genomic DNA, not chromasomal from the tested strains. The extracted DNA containing excellent overall quality and high molecular weight can be directly used for molecular biology experiments. The molecular phylogenetic dendrogram indicates that the strains used in this research were all Arthrospira platensis, which were significantly different from strains of Spirulina in molecular classification and identification in cyanobacterial genus. |
| Key words: Arthrospira platensis, whole genome DNA extraction, PCR, 16S rRNA-ITS |
