Molecular analysis of baculovirus origins of replication: identification of essential DNA sequences (replicators) and protein factors (initiators)

杆状病毒复制起点的分子分析：鉴定必需的 DNA 序列（复制子）和蛋白质因子（起始子）

• 批准号: 172834-2011
• 负责人: Carstens, Eric
• 金额: $ 2.91万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=476336
• 关键词: Molecular; analysis; baculovirus; origins; replication

Baculoviruses are distinguished by their prolific but restricted multiplication in insects. These invertebrate pathogens are used as vectors for foreign gene expression, potent biological insecticides, and gene transfer vehicles. In Canada, baculoviruses are recognized as safe biological pesticides in integrated pest management programs against economically important pests in agriculture and forestry. Some of our efforts focus on a virus that kills the Spruce budworm, the major Canadian tree defoliator. Baculoviruses have evolved multiplication strategies that limit their replication to invertebrates, but little is known of the critical factors that regulate this strict host specificity. DNA replication is one of the essential processes that likely determines host specificity. The long term goal of my research program is to identify components of the baculovirus replication system that are determinants of virus-host specificity and to use this knowledge to understand the basis for their extremely narrow host range. In an integrated series of experiments using biochemical, genetic, and cell biology approaches, the molecular mechanisms by which early genes of a model baculovirus (AcMNPV) initiate and continue DNA replication will be investigated. An obvious question is where the start points (origins) of replication are located on the AcMNPV genome. Three innovative complementary approaches will be used to investigate this question. Collectively, these studies will provide important insight into the highly conserved pathways of DNA replication that contribute significantly to the baculovirus pathogenicity. The molecular mechanisms by which baculoviruses regulate DNA replication are directly relevant to the use of baculoviruses as safe genetic vectors and biological insecticides. Ultimately, the use of baculoviruses as safe, target-specific insecticides will provide benefits to Canada's environmental sustainability and health status by reducing reliance on chemical pesticides. My program will provide stimulating and challenging projects for high quality training of personnel in an exciting area of natural sciences, insect virology. Importantly, these skills will be applicable and transferable to a multitude of career pathways.

杆状病毒的特征在于其在昆虫中的多产但受限的增殖。这些无脊椎动物病原体被用作外源基因表达的载体、有效的生物杀虫剂和基因转移载体。在加拿大，杆状病毒被公认为是防治农业和林业重要经济害虫的综合害虫管理计划中的安全生物农药。我们的一些努力集中在一种病毒，杀死云杉芽虫，加拿大主要的树木落叶。杆状病毒已经进化出限制其在无脊椎动物中复制的增殖策略，但对调节这种严格的宿主特异性的关键因素知之甚少。DNA复制是可能决定宿主特异性的重要过程之一。我的研究项目的长期目标是确定杆状病毒复制系统的组成部分，这些组成部分是病毒-宿主特异性的决定因素，并利用这些知识来了解其极窄宿主范围的基础。在一个综合系列的实验中，使用生物化学，遗传学和细胞生物学的方法，早期基因的模型杆状病毒（AcMNPV）启动和继续DNA复制的分子机制进行了研究。一个明显的问题是复制的起始点（起点）位于AcMNPV基因组上。将采用三种创新的互补办法来研究这一问题。总的来说，这些研究将提供重要的洞察高度保守的DNA复制途径，大大有助于杆状病毒的致病性。杆状病毒调控DNA复制的分子机制直接关系到杆状病毒作为安全的遗传载体和生物杀虫剂的应用。最终，使用杆状病毒作为安全的、针对特定目标的杀虫剂将通过减少对化学杀虫剂的依赖，为加拿大的环境可持续性和健康状况带来好处。我的计划将提供刺激和具有挑战性的项目，在自然科学，昆虫病毒学的一个令人兴奋的领域的人员的高质量培训。重要的是，这些技能将适用于并可转移到多种职业途径。