Expression of Vectors Transforming a Chlorella-Like Green Alga

转化类小球藻绿藻的载体表达

• 批准号: 8712041
• 负责人: Russel Meints
• 金额: $ 7.11万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1988-08-01
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8712041&HistoricalAwards=false
• 关键词: Expression; Vectors; Transforming; Chlorella; Like

Exsymbiotic, Chlorella-like eukaryotic green algae act as hosts for a group of large double-stranded DNA-containing viruses. The algae can be readily cultured in the presence of light. The viruses synchronously infect, rapidly replicate, and ultimately lyse their algal host. As a result of the development of a plaque assay, this is the first and, thus far, only example of a plant-virus system which lends itself to investigation using established technology adapted from bacteriophage studies. An additional benefit of this system is that the viral-algal lysate is a rich source of cell wall degrading enzyme activity which can be used to produce protoplasts into which recombinant DNA may be introduced. The ultimate goal of this project is to develop an algal transformation system. Toward this end, it is proposed to: (1) develop a protoplast regeneration system for this alga; (2) search for DNA sequences from either the virus or the host which will confer autonomous replication of plasmids in this alga; (3) determine the intracellular sites of viral DNA replication; (4) study viral DNA for clues as to the regulation of viral gene expression; (5) study vectors containing reporter molecules, to measure transient expression; and (6) construct vectors with selectable markers. Success in these goals will allow the characterization and exploitation of this interesting algal system, and will have significant impact on plant genetics and cell biology.

外共生的，类似小球藻的真核绿色藻类作为 一组含有大双链DNA的宿主 病毒 藻类可以容易地在以下物质存在下培养： 光 病毒同步感染，快速复制， 最终溶解藻类宿主 由于发展 这是第一个也是迄今为止唯一的一个 一个植物病毒系统， 改编自噬菌体研究的成熟技术。 一个 该系统的另外的好处是病毒-藻类溶胞产物 是细胞壁降解酶活性丰富来源， 用于产生原生质体，重组DNA可以被导入其中， 介绍 该项目的最终目标是开发藻类 转换系统 为此，建议：（1） 建立了该菌株的原生质体再生体系;（2） 寻找病毒或宿主的DNA序列， 将赋予质粒自主复制的能力;（3） 确定病毒DNA复制的细胞内位点;（4） 研究病毒DNA以寻找病毒基因调控的线索 （5）研究含有报告分子的载体， 测量瞬时表达;以及（6）构建具有 选择标记 这些目标的成功将使 这种有趣的藻类的特征和开发 系统，并将对植物遗传学和 细胞生物学