- 期刊
Karyotyping of Brassica oleracea L. Based on Cot-1 and Ribosomal DNAs
甘藍基於Cot-1 DNA及rDNA的核型分析
摘要
為了探索一種簡單、可靠、有效的甘藍核型分析新方法，我們用rDNA結合Cot-1 DNA作為甘藍染色體識別標記，成功地對甘藍進行了準確的核型分析。甘藍基因組DNA用生物素切刻平移試劑盒標記後作探針，原位雜交到有絲分裂中期相，每對染色體均顯示出特定的螢光帶型。25S及5S rDNAs分別用毛地黃素及生物素切刻平移試劑盒標記後作探針，各自原位雜交到有絲分裂中期相，有2對染色體檢測出25S rDNA雜交信號，只有1對染色體檢測出較弱的5S rDNA雜交信號。Cot-1 DNA包含rDNA，雙色螢光原位雜交顯示，25S rDNA與Cot-1 DNA雜交信號位置是一致的，表明基於rDNA及Cot-1 DNA的核型分析技術優於單獨使用一種標記技術。基於rDNA位置及Cot-1 DNA螢光帶，本研究對甘藍進行了精確的核型分析。
並列摘要
To explore a simple, reliable, and effective method of karyotyping Brassica oleracea L., Cot-1 DNA was isolated from its genome, labeled as probe with a Biotin-Nick Translation Mix kit, and in situ hybridized to mitotic spreads. Specific fluorescent bands appeared on each chromosome pair. 25S and 5S rDNAs were labeled as probes with a DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit, respectively, and in situ hybridized to mitotic preparations. Signals could be detected on two chromosome pairs for 25S rDNA, and on only one for 5S rDNA. Cot-1 DNA contains rDNA. The site identity of Cot-1 DNA and 25S rDNA on the chromosome was determined by dual-colour fluorescence in situ hybridization (FISH). It showed that the karyotyping technique based on a combination of rDNA and Cot-1 DNA chromosome markers is a superior alternative. A more exact karyotype of B. oleracea has been developed based on rDNA locations and Cot-1 DNA fluorescent bands.
延伸閱讀
- 許家言、曾夢蛟、張武男（1992）。甘藍原生質體培養與再生之研究。中國園藝，38(3)，128-138。https://doi.org/10.6964/JCSHS.199209.0128
- 陳裕星、曾勝雄、沈勳（2001）。食用紅甘蔗核糖體基因內轉錄間隔區之選殖與分析。臺中區農業改良場研究彙報，(73)，65-77。https://doi.org/10.6956/BTDAIS.200112.0065
- Huang, G. J., Huang, S. S., Chen, H. J., Chang, Y. S., Chang, S. J., Chang, H. Y., Hsieh, P. C., Chang, M. J., Lin, Y. C., & Lin, Y. H. (2009). 甘藷塊根天門冬胺酸蛋白酶cDNA之選殖和表現. Botanical Studies, 50(2), 149-158. https://www.airitilibrary.com/Article/Detail?DocID=1817406X-200904-50-2-149-158-a
- 包偉伯（2006）。甘藷葉部澱粉分支酶基因之概日韻律表現及啟動子序列分析〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU.2006.01296
- 陳鵬仁（2007）。利用雙價抗蟲基因轉殖甘藍與建立無篩選標誌系統之研究〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU.2007.01352