CRISPR/Cas9基因转录激活技术的建立及其应用

建立基于CRISPR/Cas9的基因转录激活技术,实现在HEK293细胞中激活cGAS基因的转录。首先利用慢病毒载体构建稳定表达Cas9的HEK293细胞;并采用Real-time PCR和western blot方法鉴定Cas9蛋白质表达效果。再选取不同靶序列,分别构建用于激活cGAS基因转录的sgRNA载体,将构建好的序列载体与CRISPR体系工具质粒共转染至稳定表达Cas9的HEK293细胞中。采用Real-time PCR方法检测细胞中内源cGAS的转录激活效果。成功建立了基于CRISPR/Cas9的基因转录激活技术,并在c GAS基因阴性表达的细胞中激活基因的转录。     

Genome editing and animal models

Transgenic technology allows a gene of interest to be introduced into the genome of a laboratory animal, and provides an extremely powerful tool to dissect the molecular mechanisms of disease. Transgenic mouse models made by microinjection of DNA into zygotic pro- nuclei in particular have been widely used by the genetics community for 30 years. However, it remains a rather crude approach： injected sequences randomly insert in multiple copies as concatamers, they can be mutagenic, and they have variable or silenced expression depending on the site of integration, a phenomenon called position effects. As a result, multiple lines are required in order to confirm appropriate transgene expression. This can be partially overcome by flanking transgenes with insulator sequences to protect the transgene from the influence of the sur- rounding regulatory elements. Large （〈300 kb） BAC- based transgenic vectors have also been shown to be more resistant to position effects. However, animals carrying extra copies of fairly large regions of the genome could have unpredictable phenotypes. The most effective method used to control for position effects is to target transgene insertion to specific genomic loci, the so-called targeted transgenesis; for instance, the fast, site-specific transgenic technology TargattTM. The purpose of this review is to provide an overview on the current existing methods for making targeted transgenic mouse models.     

虚拟项目教学法在模拟电子技术教学中的应用

虚拟项目教学法是基于计算机模拟仿真技术的一种教学活动。将虚拟项目教学法应用到模拟电子技术的教学中,通过项目任务的设计、项目准备、仿真实施及总体评价展开教学。该方法不仅可以达到传统项目教学法的教学目标,而且可以扩大学生的参与面,降低教学成本。实践表明,该方法适合于应用型人才培养目标下的模拟电子技术教学。     

Multisim 9在电子线路课程设计的应用

结合电子线路课程教学、电子线路课程设计实践,通过二级放大电路实例介绍Multisim 9软件在电子线路课程设计中的应用,该软件能优化电路设计,实现电路功能仿真及提高电路设计开发水平与分析电路的能力。     

Hélène Metzger: the history of science between the study of mentalities and total history

In this article, I examine the historiographical ideas of the historian of chemistry Hélène Metzger (1886–1944) against the background of the ideas of the members of the groups and institutions in which she worked, including Alexandre Koyré, Gaston Bachelard, Abel Rey, Henri Berr and Lucien Febrve. This article is on two interdependent levels: that of particular institutions and groups in which she worked (the Centre de Synthèse, the International Committee for History of Science, the Institut d'Histoire des Sciences et Techniques (Sorbonne) and the École Pratique des Hautes Études) and that of historiographical ideas. I individuate two particular theoretical aspirations pursued by the historians in Metzger's milieu: the ideal of total history and the study of the human mind. These two objectives were seen by Metzger and many others as implicating each other. Moreover, Metzger and other historians wanted to integrate the practice of commentary of texts in the realisations of those ideals. I argue, however, that these objectives proved very difficult to realise at the same time. One tradition which stemmed out of these discussions, exemplified by Bachelard, Canguilhem and Foucault, focused on the mind and knowledge, and renounced commentary of texts and total history as it was understood by the historians of the Centre de Synthèse. The latter, however, did not really pursue the study of the mind. Moreover, historians like Metzger and Koyré who practised an attentive analysis of texts could not realise total history.     

智能小车辅助平台的设计与实现

讨论了一种以教学为目的的智能小车开发平台的设计思想,描述了智能小车的机械平台结构,给出了以FreescaleMC9S08GB60为主控芯片的、开放的硬件平台设计,提供了可供用户自行“定制开发设计”的、简单易学的PC方软件平台设计。该辅助开发平台主要针对中小学课外科技活动开发,也可供高校学生进行智能小车设计、参加智能小车设计竞赛之用。     

四种操作系统共存于同一台计算机中的实现

针对很多计算机应用场合 ,要求一台计算机具有多种职能 ,需要多种操作系统支持的需求 ,本文讨论了多种操作系统共存于一台计算机中的技术     

Analysis of intron sequence variability of the conservative HMG-box of Sox9 genes in allotetraploids and their original parents

The Sox genes of allotetraploids and their original maternal red crucian carp (Carassius caassius red var.) and original paternal common carp (Cyprinus carpio L.) were detected by PCR with the designed primers based on the conserved HMG-box sequence in different species. Sequencing of Sox genes indicated that two Sox9 genes (Atsox9a and Atsox9b) existed in allotetraploids, while only one Sox9 gene existed in red crucian carp (Rcsox9a) and common carp (Ccsox9b) . All of the four Sox9 genes contained an intron in the HMG-box. with the sizes of 413 bp, 703 bp, 401 bp and 714 bp, respectively. Moreover, the introns obeyed the rule of "GT-AG" . A high similarity was observed between introns of Atsox9a and Rcsox9a (94.4%), Atsox9b and Ccsox9b (97.8%). Interestingly, the deduced amino acid sequences of their corresponding exons all shared 100% identity. Thus, introns of the HMG-domain of Sox9s in allotetraploids and their original parents have not only the length polymorphism but also intron variability. Our results provide significant molecular evidence for the origin and evolution of allotetraploids.     

基于LPC2214的Flash存储器的应用

介绍了Flash存储器K9F5608U0C和SST39VF160的特性,给出了微处理器LPC2214与它们的硬件连接电路图,以及对它们进行读、写、擦除等操作的应用软件.     

李益首次从军时间地点考辨

本文纠正了长期以来人们对李益首次从军时间和地点的错误认识,认为其首次从军时间应在大历九年至大历十二年间,而非建中初;地点在凤翔节度使李抱玉军中,而非邠宁节度使韩游瑰、凤翔节度使邢君牙或渭北节度使臧希让军中.