Upadhyay S N
Radiation Biology Department, Institute of Nuclear Medicine & Allied Sciences, Delhi.
Indian J Biochem Biophys. 2001 Dec;38(6):406-11.
Three methods, namely, absorbance of colour by reaction with Folin-Ciocalteau reagent, UV absorbance and fluorescence intensity measurements for detection of H3 histone in 0.15 M standard saline citrate (SSC) solution were compared. Maximum sensitivity was found with the Folin-Ciocalteau method. Effect of varying pH and of gamma- radiation on H3 histone and on interaction of H3 histone with DNA were studied. For this, solutions of H3 histone in SSC, in 0.9% NaCl, H3 histone + DNA in 0.9% NaCl were subjected to varying pH (1-10) and gamma- radiation (dose 10-50 Gy) and lambda(max) and Alambda(max) were monitored. From the molar ratios of histone and DNA in the complex, it was observed that at gamma -radiation dose of 50 Gy and pH 8.54, there was a depletion of 6-8 microg/ml of histone from the histone-DNA complex.
比较了三种检测0.15M标准柠檬酸盐缓冲液(SSC)中H3组蛋白的方法,即与福林-酚试剂反应的颜色吸光度、紫外吸光度和荧光强度测量法。发现福林-酚法具有最高的灵敏度。研究了不同pH值和γ辐射对H3组蛋白以及H3组蛋白与DNA相互作用的影响。为此,将SSC中、0.9%NaCl中的H3组蛋白溶液以及0.9%NaCl中的H3组蛋白+DNA溶液分别置于不同pH值(1-10)和γ辐射(剂量10-50Gy)下,并监测λ(max)和Δλ(max)。从复合物中组蛋白与DNA的摩尔比可以看出,在γ辐射剂量为50Gy且pH值为8.54时,组蛋白-DNA复合物中的组蛋白减少了6-8μg/ml。