组蛋白-DNA共振光散射法的建立及应用

杜晓燕; 郑晶; 王保珍

组蛋白-DNA共振光散射法的建立及应用

Study on resonance light-scattering of histene-DNA and its application

摘要

摘要:
目的建立灵敏度高、简单方便、试剂安全的共振光散射方法测定生物体DNA含量。
方法在pH5.5的HAc-NaAc缓冲体系中, 将组蛋白与DNA混合, 在入射光波长和荧光波长相同条件下, 用普通荧光分光光度计测定溶液的共振光散射强度; 光散射强度与DNA浓度呈良好线性关系时测定核酸含量; 应用该法检测鼠肝和酵母体内的DNA含量。
结果扫描光谱显示最大光散射波长为551nm, 测定最佳条件为pH5.5; 组蛋白用量10μg/ml。ctD-NA、fsDNA、hpDNA等3种核酸均与光散射强度有良好线性关系, 相关系数均大于0.99。与常规分光光度法对照测定样品DNA含量, 相对误差 < 10%。
结论该方法灵敏度优于常规方法2~3个数量级, 快速方便, 适合于生物体内核酸含量测定。

Abstract:
Objective To establish a sensitive, simple and safe method for the determination of nucleic acid in living body.
Methods In the acetic acid-sodium acetate buffer solution at pH5.5, on the basis of the measarement of the light-scattering from a complex formed by DNA and histone using a common spectrofluorometer, the DNA in mouse liver and yeast was measured.
Results The optimum measurement for DNA was at the wavelength of 551 nm; pH5.5;histon of 10μg/ml.The linear relationship between the light-scattering intensity(IRLS)and the concentration of nucleic acids was good(r > 0.99).The agreement between the light-scattering measurement and the conventional spectrophotometric method was satisfactory(relative error < 10%).
Conclusion The method was more sensitive than conventional spectrophotometic method, qaick, convenient and reliable with little interference.

HTML全文

参考文献(5)

施引文献

资源附件(0)