【摘要】 目的:为了建立细菌DNA G+Cmol%的快速准确的测定方法,选用反相高效液相色谱法测定Bacillus claussiDNA的G+C mol%。方法:以已完成全基因测序的Bacillus haloduransC-125作为标准菌株,作者实验室新分离鉴定的2株Bacillus claussi作为待测菌株。采用80%20mmol/L磷酸二氢钾溶液(pH5.6)20%甲醇为移动相,检测波长260nm,流速1ml.min-1,在Kromasil C18柱上对4种碱基进行分离。结果:DNA碱基分离效果好,无杂峰干扰,以峰面积计算得到标准菌株Bacillus haloduransC-125的G+C mol%为44.23%,待测菌株的G+C mol%分别为45.75%、43.35%,经过统计学分析,与已经报道的G+C mol%差异不显著。结论:实验结果充分显示高效反相液相色谱法测定细菌DNAG+C mol%快速、准确、结果稳定,是细菌精确分类鉴定的一种可靠方法。更多还原

【Abstract】 Objective:To determinate two recently isolated Bacillus claussi strains′DNA G+Cmol% rapidly and accurately,reversed-phase high performance liquid Chromatography was used.Methods:The full-sequenced Bacillus halodurans C-125 was selected as standard strain.Various bases were separated on a Kromasil C18 column by using 80% KH₂PO₄ buffer(20mmol·l^-1,pH5.6) and 20% methanol as mobile phase at flow rate of 1ml·min^-1.The UW detection was set at 260nm.Results:The G+C mol% of Bacillus halodurans C-125 and the two isolated strains were 44.23%,45.75%,43.35% separately,which were closely match to the G+C mol% of the documental values after the analyze by Statistics.Conclusion:The experimental results demonstrate that this method is rapid,accurate and reliable for bacterial classification.更多还原