Investigating the Rex exclusion genes of bacteriophage Lambda

噬菌体生物学已经成为我们目前理解分子遗传学和设计新的分子生物学工具的关键门户。虽然在这一领域的工作已经导致了对基因调控，DNA复制和重组的突破性理解，但仍有许多东西有待学习，许多具有潜在可利用利益的谜团仍未解决。其中一个特别重要的是λ T4 rII排除（雷克斯）表型。这种表型是一种复杂的、神秘的现象，被定义为噬菌体T4快速裂解突变体（称为rII突变体）不能在大肠杆菌的细胞菌苔上形成噬斑。大肠杆菌携带休眠的λ噬菌体，整合在E. coli染色体。具有讽刺意味的是，尽管rII基因在逃避雷克斯表型方面具有历史意义，并且是迄今为止最彻底的定位基因，但雷克斯的RII逃避机制和RII排斥表型本身仍然无法解释。这种表型的阐明可能会导致我们走向类似的真核病毒排斥系统，可能会利用在新的抗病毒策略的发展。该提案旨在确保关键人员和设备进行这些遗传和生物化学测定，以寻求显着扩大我们的理解和巩固与λ雷克斯基因（赋予雷克斯）相关的表型和rII基因抑制这种强大的排斥表型的机制。这里描述的一些实验已经证明了对Lambda T4 rII排除机制的阐明的初步成功。这里概述的研究不仅有助于促进我们对经典遗传之谜的理解，并提供有关这个神秘系统的重要科学信息，更重要的是，翻译这些发现-识别可能用于开发新型抗病毒药物的类似人类病毒编码的排除系统。拟议的计划还提供了一个坚实的科学平台，培养高素质的人才，在分子遗传学，噬菌体生物学和核酸和蛋白质生物化学的综合专业知识。

Bacteriophage biology has served as critical gateway to our current understanding of molecular genetics, and to the design of new molecular biology tools. Although work in this area has led to ground breaking understanding of gene regulation, DNA replication and recombination, much remains to be learned and many mysteries with potentially exploitable benefits remain unsolved. One of particular significance is the Lambda T4rII exclusion (Rex) Phenotype. This phenotype is a complex, enigmatic phenomenon that is defined as the inability of a bacteriophage T4 rapid lysis mutant (called rII mutant), to form plaques on a cell lawn of E. coli carrying a dormant Lambda bacteriophage, integrated in the E. coli chromosome. Ironically, despite the historical significance of the rII genes in evading the Rex phenotype, and being the most thoroughly mapped genes of any to date, the mechanisms of RII evasion of Rex and the RII exclusion phenotype itself, remain unexplained. The elucidation of this phenotype may lead us toward similar eukaryotic virus exclusion systems that may be exploited in the development of new antiviral strategies. This proposal seeks to secure critical personnel and equipment to carry out these genetic and biochemical assays that seek to significantly extend our understanding and consolidate the phenotypes associated with the Lambda rex genes (that confer Rex) and the mechanism by which the rII genes suppress this powerful exclusion phenotype. Some of the experiments described here have proven preliminary success toward the elucidation of the Lambda T4rII exclusion mechanism. The research outlined here serves to not only advance our understanding of a classical genetic mystery and provide important scientific information about this enigmatic system, but more importantly, translate these findings--identifying similar human virus-encoded exclusion systems that may be exploited for development of novel antivirals. The proposed program also provides a solid scientific platform to develop highly qualified personnel with combined expertise in molecular genetics, phage biology and nucleic acid and protein biochemistry.