Bacteriophage lambda replication and phage display

噬菌体 lambda 复制和噬菌体展示

• 批准号: 138296-2012
• 负责人: Hayes, Sidney
• 金额: $ 1.89万
• 依托单位: University of Saskatchewan
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=505684
• 关键词: Bacteriophage; lambda; replication; phage; display

Our research program involves fundamental studies on bacteriophage lambda virus replication, and genetic engineering attempts to develop a platform for making multiplication-defective lambda virus particles displaying gene fusions. These viral particles can have utility as vaccines and for gene therapy. The platform accomplishes the divergent tasks of enabling very high concentration production of the viral particles, yet effectively killing the virus by introducing genetic modifications that prevent it from multiplying in animal cells or in the environment. The platform will simultaneously produce encapsulated viral DNA and gene-fusion display particles. The gene-fusions displayed on the viral particles enable them to recognize target proteins on specific animal cells, enhancing recognition and viral uptake. Once in the cell, the gene expression cassette that was inserted into the viral genome will encode for the synthesis of a protein(s) that can induce a vaccine response, or support gene therapy. Dr. Hayes has successfully designed gene fusion phage display vaccines to PorcinePCV2 and Bovine BVDV2 viruses, to Ebola and Marburg viruses, and several to anthrax. Advantages of symbiotic research goals: Many of the platform technology gene expression vectors have proven of immense value to our gene expression and viral replication studies. Here, we try to understand how replication initiation from the lambda virus is silenced, which we found involves the participation of a micro RNA. We have identified lambda mutations that escape silencing (Sip) and seek to characterize their effect. Studies also focus on the activities of the lambda replication initiation genes O and P, and host cell proteins. Our particular focus is the interaction between the host DnaB helicase required to unwind DNA during replication and P, which is very toxic to the cell, and appears to stimulate a very high frequency of chromosomal mutation. Lambda escapes the effects of P. We seek to characterize the effects of P on host cell metabolism, and the regulation of gene P expression and P activity in cell.

我们的研究计划涉及噬菌体λ病毒复制的基础研究，基因工程试图开发一个平台，使增殖缺陷λ病毒颗粒显示基因融合。 这些病毒颗粒可以用作疫苗和基因治疗。 该平台完成了不同的任务，即能够非常高浓度地生产病毒颗粒，同时通过引入遗传修饰来有效地杀死病毒，从而防止病毒在动物细胞或环境中繁殖。 该平台将同时产生封装的病毒DNA和基因融合展示颗粒。 病毒颗粒上显示的基因融合使它们能够识别特定动物细胞上的靶蛋白，增强识别和病毒摄取。 一旦进入细胞，插入病毒基因组的基因表达盒将编码合成可以诱导疫苗应答或支持基因治疗的蛋白质。 Hayes博士成功设计了针对猪PCV 2和牛BVDV 2病毒、埃博拉病毒和马尔堡病毒以及几种炭疽的基因融合噬菌体展示疫苗。 共生研究目标的优势：许多平台技术基因表达载体已被证明对我们的基因表达和病毒复制研究具有巨大价值。 在这里，我们试图了解从λ病毒的复制启动是如何沉默的，我们发现这涉及到一个微小RNA的参与。 我们已经鉴定了逃避沉默（Sip）的λ突变，并试图表征它们的作用。研究还集中在λ复制起始基因O和P以及宿主细胞蛋白的活性上。 我们特别关注的是宿主DnaB解旋酶之间的相互作用，所需的DNA在复制和P，这是非常有毒的细胞，并出现刺激一个非常高的频率的染色体突变。 我们试图描述P对宿主细胞代谢的影响，以及细胞中P基因表达和P活性的调节。