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: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=593396
• 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.

我们的研究项目涉及噬菌体 lambda 病毒复制的基础研究，以及基因工程尝试开发一个平台，用于制造显示基因融合的增殖缺陷 lambda 病毒颗粒。这些病毒颗粒可用作疫苗和基因治疗。该平台完成了不同的任务，即能够以非常高的浓度生产病毒颗粒，同时通过引入阻止病毒在动物细胞或环境中繁殖的基因修饰来有效杀死病毒。该平台将同时生产封装的病毒 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目标蛋白，从而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诱导疫苗反应或支持基因治疗的蛋白质。 Hayes 博士成功设计了针对猪 PCV2 和牛 BVDV2 病毒、埃博拉病毒和马尔堡病毒以及多种炭疽病毒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 lambda 病毒的复制起始是如何被沉默的，我们发现这涉及到微小 RNA 的参与。我们已经确定了逃避沉默 (Sip) 的 lambda 突变，并试图表征其效应。研究还集中于 lambda 复制起始基因 O 和 P 以及宿主细胞蛋白的活性。我们特别关注复制过程中解旋 DNA 所需的宿主 DnaB 解旋酶与 P 之间的相互作用，P 对细胞具有很强的毒性，并且似乎会刺激非常高频率的染色体突变。 Lambda 逃避 P 的影响。我们试图表征 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。