Kunnath L, Locker J
Nucleic Acids Res. 1985 Jan 11;13(1):115-29. doi: 10.1093/nar/13.1.115.
We have analyzed the DNaseI sensitivity of chromatin from the rat albumin and alpha-fetoprotein genes in the fetal liver (which synthesizes albumin and alpha-fetoprotein), adult liver (which synthesizes albumin), fetal yolk sac (which synthesizes alpha-fetoprotein), and adult kidney (which synthesizes neither). Active genes were much more sensitive than their kidney counterparts, and the adult liver alpha-fetoprotein and fetal yolk sac albumin genes showed intermediate levels of sensitivity. Sensitivity was analyzed as a function of the extent of DNaseI digestion. Rate constants were calculated for the degradation of individual DNA hybridization bands and normalized to the intrinsic rate constants of the same bands degraded in purified DNA. This enabled us to eliminate the inconsistencies that otherwise result from comparing chromatin sensitivity of different DNA sequences, or chromatin sensitivity in different nuclear environments.
我们分析了来自胎肝(合成白蛋白和甲胎蛋白)、成体肝脏(合成白蛋白)、胎黄囊(合成甲胎蛋白)和成年肾脏(两者均不合成)的大鼠白蛋白基因和甲胎蛋白基因染色质的脱氧核糖核酸酶I(DNaseI)敏感性。活性基因比它们在肾脏中的对应基因更敏感,而成体肝脏甲胎蛋白基因和胎黄囊白蛋白基因显示出中等水平的敏感性。敏感性分析是作为DNaseI消化程度的函数进行的。计算了各个DNA杂交条带降解的速率常数,并将其标准化为在纯化DNA中降解的相同条带的固有速率常数。这使我们能够消除因比较不同DNA序列的染色质敏感性或不同核环境中的染色质敏感性而产生的不一致性。