STUDY ON THE ANALYSIS OF TAXONOMIC DISTANCE AMONG

植物分类距离分析研究

基本信息

批准号：
09557024
负责人：
EZAKI Takayuki
金额：
$ 6.08万
依托单位：
GIFU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557024/
关键词：
taxonomy species definition DNA hybrid ribosome Phylogeny Liquid phase hybridization リボソーム 16S リボゾーム DNA 相同性種

项目摘要

Prokaryotes were phylogenetically classified by 16SrRNA sequences and genetic distance among independent species were quantitatively measured simply by comparing their 16SrRNA differences. However, it become clear that some established species shares same 16SrRNA sequences.In this case, we need to use other method to differentiate closely related species. Quantitative DNA/DNA hybridization method is a officially accepted method to define a bacterial species. However, this method is difficult to perform and DNA used for this method is often cotaminated with polysaccharide, which interfers quantitative analysis of DNA/DNA hybrid. We established several parameters to evaluate DNA purity and a rapid measuring method of guanine plus cytosine contents of DNA to define DNA/DNA hybridization experiments. RNA and polysaccharide contamination was successfully predicted by measuring absorbance of 235/260 nm and Anthrone quantitation method of sugars. Guanine plus cytosine content of DNA was instantly measured by Light cycler with SYBR Green 1 fluorescence. This information is very useful to define quantitative DNA/DNA hybridization at optimal temperature. Trials to differentiate strains within a species were successfully introduced by many scientists. We used Ceul restriction enzyme to evaluate distances among strains. This method has a advantage because bands generated by the Ceul digestion indicate numbers of copies of ribosomal RNA operon. From fragment analysis of generated size offered useful information to analyze gene recombination history.

原核生物通过16sRRNA序列进行了系统发育分类，而独立物种之间的遗传距离仅通过比较它们的16sRRNA差异来定量测量。但是，很明显，某些已建立的物种具有相同的16sRRNA序列。在这种情况下，我们需要使用其他方法来区分密切相关的物种。定量DNA/DNA杂交方法是定义细菌物种的官方接受方法。然而，这种方法难以执行，用于该方法的DNA通常用多糖进行摄影，该多糖将DNA/DNA杂交的定量分析互动。我们建立了几个参数来评估DNA纯度和DNA的鸟嘌呤和胞嘧啶含量的快速测量方法，以定义DNA/DNA杂交实验。通过测量235/260 nm的吸光度和糖量定量方法，成功预测了RNA和多糖污染。通过使用SYBR绿色1荧光的光循环仪立即测量DNA的鸟嘌呤加上DNA的胞嘧啶含量。该信息对于在最佳温度下定义定量DNA/DNA杂交非常有用。许多科学家成功地引入了区分物种中菌株的试验。我们使用CEUL限制酶来评估菌株之间的距离。该方法具有优势，因为CEUL消化产生的频带表明核糖体RNA操纵子的副本数量。从碎片分析生成的大小提供了有用的信息来分析基因重组历史。