紫山药组织RNA的提取方法

针对紫山药富含次生代谢物质的特点,探索出一种可有效去除其酚类、多糖和DNA等杂质的紫山药总RNA提取方法。在SDS法的基础上,于粉碎材料时,用水不溶性聚乙烯吡咯烷酮(PVP)防止酚类物质氧化,同时以高盐溶液去除多糖污染。利用这一方法提取的紫山药多个组织的总RNA,经琼脂糖电泳鉴定,可见清晰的18和28SRNA的2条带型,表明RNA完整,无弥散或降解;经过紫外吸收分析,A260/A380、A260/A2300分别在1.86~2.01,2.17~2.65,纯度良好,所提取的总RNA质量可以满足进一步分子生物学研究的要求。     

Mechanistic aspects of the interaction of intercalating metal complexes with nucleic acids

Since the discovery of the intercalative binding mode, almost half a century ago, intense efforts have been devoted to design, synthesize and test new small molecules that can bind nucleic acids with improved recognition and affinity. Among them, metal bearing compounds play a principal role. Despite the plethora of different metal complexes which have been designed to react with DNA and which have been tested, the binding mechanisms have often not been analysed. This is unfortunate, considering the importance of understanding of the binding features in depth in order to optimise their biological effects. This review covers articles where an analysis of the kinetic aspects of the interaction between the target metal compound and nucleic acids has been carried out and details of the reaction mechanism are provided. Flat metal complexes (porphyrins), spherical complexes with protruding intercalating residues, azamacrocycle metallo-intercalators and intercalators with metal bearing pendant arms are the classes of molecules that have been taken into account. The limits of the SDS method, employed to measure the rates of drug dissociation from polynucleotides, are also discussed.     

反BIRD在NMR谱编辑中的应用

建议了一个新的双线性旋转去偶序列 ,称为反BIRD (inverse -BIRD) ,其去偶作用与碳连质子的数目有关。将反BIRD结合到INEPT实验中 ,得到了INEPT -iBIRD实验技术 ,它可用于核磁共振碳谱的编辑与简化     

对DEPT实验技术的一点改进

将DEPT -1 3 5实验的 1 3 5° -脉冲分成一个 90°脉冲和一个 4 5°脉冲 ,通过相位循环 ,设计了两个新的DEPT实验分别称为DEPT-90 -4 5和DEPT -90 +4 5。DEPT -90 -4 5谱中只有 -CH3的吸收峰。DEPT -90 +4 5谱中没有 >CH -的吸收峰 ,其 -CH3为正信号 ,-CH2 -为负信号。实验结果显示 ,对比DEPT -90谱 ,这两个新的DEPT谱中完全没有所选基团之外的残余干扰信号 ,而且DEPT -90 -4 5谱比DEPT-90 +4 5谱更好。     

Co-ion dependence of DNA nuclease activity suggests hydrophobic cavitation as a potential source of activation energy

The source of the activation energy that allows cutting of DNA by restriction enzymes is unclear. A systematic study of the cutting efficiency of the type-II restriction endonuclease EcoRI, with varying background electrolyte ion pair and buffer reported here, shows only a modest dependence of efficiency on cation type. Surprisingly, efficiency does depend strongly on the presumed indifferent anion of the background salt. What emerges is that competition between the background salt anion and the buffer anion for the enzyme and DNA surfaces is crucial. The results are unexpected and counterintuitive from the point of view of conventional electrolyte theory. However, taken together with recent developments in surface chemistry, the results do fall into place and could also suggest a novel mechanism for enzyme activity as an alternative to metal-activated hydrolysis: microscopic cavitation in a hydrophobic pocket might be the source of activation energy. Received 19 June 2000 and Received in final form 17 October 2000