过氧乙酸-TAE缓冲液对硫修饰DNA切割条件的优化

DNA Modification by Sulfur: Analysis of Dnd Phenotype by Peracetic Acid-TAE Treatment

用乙酸和过氧化氢反应生成的过氧乙酸(PAA)辅配三羟甲基氨基甲烷-乙酸-EDTA(TAE,pH7.5)对硫修饰DNA的切割条件进行了优化.用6 mmol/L的PAA-TAE处理硫修饰基因组和质粒DNA,30 min可使降解程度达到最大饱和.用60 mmol/L的PAA-TAE经20 h处理低熔点琼脂糖包埋的菌丝体DNA,得到最佳切割效果.用PAA-TAE验证了一株在脉冲场凝胶电泳过程中基因组DNA降解的Salmonella菌株的Dnd表型;同时验证其质粒激活降解与已经验证的Streptomyces lividans质粒激活降解一样为位点专化性硫修饰.

This paper tested the use of PAA combined with TAE （PAA-TAE） as a cleaving agent to analyze the Dnd phenotype of the sulfur modified chromosomal and plasmid DNA. A 6 mmol/L PAA-TAE concentration for 30 min treatment was found to be optimal. For cleavage of chromosomal DNA in plugs, the best treatment was 60 mmol/L PAA-TAE for 20 h. The Dnd phenotype of Salmonella serovar Typhimurium 87, of which the chromosomal DNA was also found degraded during pulse field gel electrophoresis when analyzed by PAA-TAE treatment, was reproducibly obtained.