In vivo study of gene regulation and gene function in Sulfolobus ssp.: application and development of transformation vectors

硫化叶菌基因调控和基因功能的体内研究：转化载体的应用和开发

• 批准号: 5403955
• 负责人: Privatdozent Dr. Georg Lipps
• 金额: --
• 依托单位: Hochschule für Life Sciences FHNW
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5403955?language=en
• 关键词: vivo; study; gene; regulation; function

Sulfolobus represents one of the best studied hyperthermophilic archaea of the kingdom crenarchaeota and has developed into an important model organism in particular for the understanding of information processing (i.e. replication, transcription, translation) in archaea. However, no efficient genetic system is yet available for Sulfolobus and for other hyperthermophiles. Recent efforts in our laboratories including the characterization of a Sulfolobus plasmid and the development of a stable and selectable vector system have opened up new avenues to study gene regulation and protein/protein interaction in vivo and to refine and to develop genetic tools. We will use our novel vector system for the characterization of regulated promoters in vivo, and for the expression of tagged proteins. Furthermore, we will refine the transformation system such that it will allow high efficient cloning in Sulfolobus and we want to develop novel tools for mutagenesis and/or gene silencing that will allow the straight forward study of gene functions in a hyperthermophilic archaeon. The proposed approaches include the development of novel markers and DNA uptake techniques with liposomes, a helper phage system, transposon mutagenesis and gene silencing by homologous recombination or RNA interference.

Sulfolobus代表了王国Crenarchaeota的最富有疗法的过度古细菌之一，并已发展成为一个重要的模型生物体，特别是在古细菌中理解信息处理（即复制，转录，翻译）。但是，尚无有效的遗传系统可用于磺胺子和其他热疗。我们的实验室的最新努力，包括表征磺胺质质粒的表征以及稳定且可选的矢量系统的发展，已经开辟了新的途径，以研究体内基因调节和蛋白质/蛋白质相互作用，并改进并开发基因工具。我们将使用新型的矢量系统来表征体内调节启动子的表征，以及标记蛋白的表达。此外，我们将完善转化系统，使其将允许在磺bo子中进行高效的克隆，并且我们希望开发新的工具进行诱变和/或基因沉默，从而可以直接研究高疗法中的基因功能。提出的方法包括使用脂质体的新标记和DNA摄取技术的发展，辅助噬菌体系统，转座子诱变以及通过同源重组或RNA干扰的基因沉默。