- 學位論文
以毛細管電泳法(CE)和變性凝膠電泳法(DGGE)分析環境中抗卡那黴素(Kanamycin)細菌分佈之可行性研究
Using Capillary Electrophoresis(CE) and Denaturing Gradient Gel Electrophoresis (DGGE) for Determing the Distribution of Bacteria Resistant to Kanamycin
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摘要
近年來由於環境改變，土壤微生物多受到了干擾，故探討土壤菌相改變為重要的課題。早期相關技術如：RAPD、RFLP、DGGE、RISA…等已被運用在菌相分析上，上述技術多以16S rDNA為主，近年以16S-23S（900bp~1100bp） rDNA(ITS，0.5-1.4Kb)進行之相關研究也有增加的趨勢。本研究利用培養法(選擇性培養基LB，LB＋Kan＋Cyc)與化學法(CE和DGGE)進行種植基因改造木瓜隔離試驗田對土壤中一般細菌與KanR菌相分佈的分析。結果顯示利用DGGE技術配合16S rDNA中V3區域進行土壤中總DNA之一般細菌菌相分析，發現不同土壤樣本總DNA呈現不同的DGGE圖譜，經叢聚分析（Cluster analysis）顯示可得到基因改造作物對一般細菌菌相分佈的差異性。利用16S rDNA（1.4Kb）及ITS配合配合BLAST比對序列也可得到不同土壤樣本中KanR細菌分佈。
並列摘要
In recent years due to environmental changes, and more by the disruption of soil microbes, the soil bacteria to explore the importance of changing the subject. Early-related technologies such as: RAPD, RFLP, DGGE, RISA ... and have been used for analysis of bacteria, many of these technologies to 16S rDNA-based in recent years to 16S-23S (900bp ~ 1100bp) rDNA (ITS, 0.5~1.4 Kb ) carried out research is also increasing. In this study, culture method (selective medium LB, LB + Kan + Cyc) and chemical methods (CE and DGGE) for the cultivation of genetically modified papaya in isolated experimental field of soil bacteria with the general distribution of bacteria KanR analysis. The results showed that with the use of DGGE technology V3 region of 16S rDNA of total DNA of soil bacteria of the general analysis of bacteria and found that the total DNA of different soil samples showed different DGGE patterns, the clustering analysis (Cluster analysis) shows available on genetically modified crops general bacterial strain differences in distribution. The use of 16S rDNA (1.4Kb) and with the ITS sequence comparison with BLAST is also available in the soil samples of different bacteria KanR distribution.
參考文獻
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