反相高效液相色谱法测定色盐杆菌新种ST307的DNA G+C mol%含量

Determination of the DNA G+ C mol% in Type Strain ST307 of Prospectively New Species of Genus Chromohalobacter by Reversed-phase High Performance Liquid Chromatography

目的:使用反相高效液相色谱法测定色盐杆菌新种ST307的DNAG+Cmol%含量。方法:以Escherichia coli DH5α为标准菌株,采用90%重蒸水10%甲醇为流动相,检测波长260nm,流速1ml.min-1,在VenusilM P C18柱上对四种碱基进行分离。结果:DNA碱基分离效果好,以外标法计算得到标准菌株DH5α的DNAG+Cmol%含量为50.3%,待测菌株ST307的DNAG+Cmol%含量为60.5%。结论:采用反向高效液相色谱法测定色盐杆菌的DNAG+Cmol%含量准确可靠。

Objective：To determinate the DNA G＋Cmol % of a novel strain ST307 of Chromohalobacter genera,reversed-phase high performance liquid chromatography was used.Method：Escherichia coli DH5α was selected as standard strain.Various bases were separated on a Venusil MP C18 column by using 90% redistilled water and 10% methanol as mobile phase at flow rate of 1ml·min-1.The UV detection was set at 260nm.Result：The G＋C mol% of DH5α and ST307 were 50.3 and 60.5 separately.Conclusion：The experimental results demonstrate that this method is accurate and reliable for bacterium of Chromohalobacter genera classification.