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

作　　者：梁晶丹[1] 汪志军[1] 周秀芬[1] 邓子新[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海200030

出　　处：《上海交通大学学报》2008年第1期5-10,共6页Journal of Shanghai Jiaotong University

基　　金：国家自然科学基金资助项目(30630001)

摘　　要：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.

关 键 词：硫修饰DNA 过氧乙酸-三羟甲基氨基甲烷-乙酸-EDTA 激活 DNA降解 

分 类 号：Q555.4[生物学—生物化学]