HPLC技术对细菌基因组DNA碱基组成的精确测定  

Precise Determination of Base Composition in Bacterial Genome DNA Using HPLC Techniques

作　　者：李越中[1] 胡玮[1] 张禹清[1] 张勇[1] 韩冠君[1] 黎志凤[1] 王冰[1] 

机构地区：[1]山东大学

出　　处：《化学学报》2001年第9期1464-1470,共7页Acta Chimica Sinica

基　　金：国家自然科学基金，高等学校博士学科点专项科研项目

摘　　要：高效液相色谱技术(HPLC)在精确分析细菌基因组DNA碱基组成时存在着重复性差和杂质干扰的问题.本研究结合反相HPLC技术对化学水解(高氯酸法和甲酸法)和生物酶水解(磷酸二酯酶I和牛肠粘膜碱性磷酸酶)DNA的水解条件和产物进行了分析,建立了一套系统的分析方法,改善了分析的重复性问题,排除了主要的误差来源,达到了对细菌DNA碱基组成精确分析测定的目的.Errors of reproducibility with high performance liquid chromatography ( HPLC) are remarkably serious in determination of base composition of bacterial genome DNA, which is usually defined as guanine plus-cytosine content [(G + C) mol% ]. For partially purified DNA, salts are a major source of interference in enzymatic degradation and RNAs are a major source of interference in chemical degradation. This research analyzes the conditions and products by chemical (perchloric acid and formic acid) and enzymatic degradations (phasphodiesterase I and bovine intestinal mucosa alkaline phophatase) of bacterial genome DNA using RP HPLC, and introduces a systematic method to precisely measure the base composition of bacterial DNA.

关 键 词：高效液相色谱 细菌 DNA 碱基组成 化学水解 酶水解 测定 基因组 

分 类 号：Q936[生物学—微生物学]