DNA甲基化检测方法的研究进展

DNA甲基化是一种重要的遗传外修饰,是表观遗传学(epigenetics)的重要组成部分[l]。它参与了动物胚胎发育、基因印迹和X染色体失活等过程,在基因表达的调控中具有重要作用,异常甲基化可以导致肿瘤的形成。近20年来,DNA甲基化的研究逐渐成为新的研究热点。随着对甲基化研究的不断深人,各种各样甲基化检测方法被开发出来以满足不同类型研究的要求。本文对目前现有常用的CpG岛甲基化检测方法作一综述,并着重介绍几项以芯片技术为基础的高通量研究DNA甲基化的方法。     

基于小波分析的水稻蛋白OsPCBP的Hurst指数的提取

用DNA walk方式将水稻蛋白OsPCBP的DNA序列映射为相应的数字信号,并利用小波变换提取其Hurst指数.结果分析表明,在OsPCBP的DNA序列中存在长程相关性.     

Tagged Probe Interval Graphs

A generalization of interval graph is introduced for cosmid contig mapping of DNA. A graph is a tagged probe interval graph if its vertex set can be partitioned into two subsets of probes and nonprobes, and a closed interval can be assigned to each vertex such that two vertices are adjacent if and only if at least one of them is a probe and one end of its corresponding interval is contained in the interval corresponding to the other vertex. We show that tagged probe interval graphs are weakly triangulated graphs, hence are perfect graphs. For a tagged probe interval graph with a given partition, we give a chordal completion that is consistent to any interval completions with respect to the same vertex partition.     

链霉菌查尔酮合成酶基因克隆及原核表达载体构建

根据Genbank数据库已知的链霉菌的查尔酮合成酶基因的保守区设计chs基因特异简并引物,土壤总DNA为模板,利用PCR技术扩增得到该l条chs基因编码区,通过TA克隆、测序和同源比对及进化分析表明:该基因为放线菌来源chs基因.分别在该基因5’末端和3’末端分别引入的限制酶NcoI和EcoR I酶切位点,利用上述2种限制酶分别酶切导入到psimple-T/chs载体和原核表达pET32a,凝胶回收目的片段后,将二者连接并转化大肠杆菌感受态细胞,转化子经菌液PCR筛选、双酶切鉴定后,3730测序结果表明,该基因全长编码区为1089 bp,推测该基因编码全长为362个氨基酸残基,等电点(PI)为5.41、分子量为3 965道尔顿含有CHS保守功能区的酸性蛋白质.分析表明,该基因与Streptomyces lividans来源的查尔酮合成酶RppA基因核苷酸相似性高达93％,氨基酸序列相似性高达87.70％.测序结果表明,该基因已经成功插入到pET32a载体中.     

多通道DNA测序仪的新型设计

目前大多数商品化的DNA测序仪的信号检测系统是基于CCD或PMT。基于CCD的系统需要对CCD进行制冷,其造价相对较高且灵敏度相对较低;基于PMT的系统使用机械扫描来实现多通道检测,其工作稳定性相对较低且工作的机械噪音大。本文提出了新型的DNA测序仪,它采用PMT共焦荧光系统,通过f-θ透镜和光学扫描来实现多通道并行检测。在此新系统中采用标准双链DNA样品:pBR322/HaeIII(使用TO染料)进行了毛细管电泳实验,测得系统的检测限可达:1.1841×10-11mol/L。新型DNA测序仪的工作机械噪声相比基于机械扫描的系统要低得多,且工作稳定性和灵敏度也很高。此系统可应用于基于激光诱导荧光的毛细管阵列和多通道微芯片的电泳检测。     

花粉管通道转导外源野生稻DNA进入栽培稻的方法研究

本文报道了在实验室(空调培养)、温室及田间等不同条件下,利用花粉管通道转导外源野生稻 DNA 进入栽培稻,并将 D_0代外源 DNA 重组体幼胚进行培养,威功地获得了一批外源野生稻 DNA 重组体植株的实验方法。结果表明,用花粉管通道转导外源 DNA 的整体系统是简单易行的方法。花粉管通道转导外源野生稻DNA 与 D_0代重组体幼胚离体培养相结合的技术是在分子水平上改良稻种资源和进行水稻分子育种的有效途径。     

Risk Assessment for Aflatoxin: I. Metabolism of Aflatoxin B1 by Different Species

A comparison of the generation and detoxification of reactive aflatoxin B1 metabolites in different species may elucidate why animal species vary widely in sensitivity to aflatoxin B1 carcinogenicity, in addition to offering some perspective on how sensitive man may be to the carcinogenic effects of this mycotoxin. Scientific literature comparing the ability of cellular fractions from different species to metabolize aflatoxin B1 is reviewed. However, in vitro studies exclude components for multiple metabolic pathways, eliminating the possibility of competition between metabolic activation and detoxification. Quantification of metabolic activation by different species from these studies is therefore limited. The species-specific carcinogenic potency of aflatoxin B1 may be reflected in levels of aflatoxin B1-DNA adducts generated in vivo. The current paucity of quantitative information on human DNA adduct levels does not permit comparisons between different species on this basis.     

组件技术和Windows DNA体系结构

主要阐述组件技术和W indows DNA技术,并分析了采用三层结构进行开发的必要性。     

反义寡核苷酸技术

本文基于反义寡核苷酸技术的基本原理，评述了该技术的主要内容和近些年来的研究进展；着重介绍了其在农业、医药和生物学基础研究等方面的应用．此外，还分析了此技术在应用方面存在的一些问题，并展示了其应用前景．     

Fitting Weibull duration models with random effects

Duration time models often should include correlated failure times, due to clustered data. These random effects hierarchical models sometimes are called frailty models when used for survival analyses. The data analyzed here involve such correlations because patient level outcomes (the times until graft failure following kidney transplantation) are observed, but patients are clustered in different transplant centers. We describe fitting such models by combining two kinds of software, one for parametric survival regression models, and the other for doing Poisson regression in a hierarchical setting. The latter is implemented by using PRIMM (Poisson Regression and Interactive Multilevel Modeling) methods and software (Christiansen & Morris, 1994a). An illustrative example for profiling data is included withk=11 kidney transplant centers andN=412 patients.