Investigating the Rex exclusion genes of bacteriophage Lambda

噬菌体生物学已经成为我们目前对分子遗传学的理解和设计新的分子生物学工具的关键门户。尽管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，许多具有潜在开发价值的奥秘仍未解决。其中一个特别重要的是Lambda T4rII排除（Rex）表型。这种表型是一种复杂而神秘的现象，它被定义为噬菌体T4快速裂解突变体（称为rII突变体）无法在大肠杆菌的细胞草坪上形成斑块，大肠杆菌染色体上携带一个休眠的Lambda噬菌体。具有讽刺意味的是，尽管rII基因在逃避Rex表型方面具有历史意义，并且是迄今为止最彻底定位的基因，但rII逃避Rex的机制和rII排除表型本身仍未得到解释。这种表型的阐明可能会引导我们走向类似的真核病毒排斥系统，这可能会在开发新的抗病毒策略中被利用。本提案旨在确保关键人员和设备来进行这些遗传和生化分析，这些分析旨在显著扩展我们的理解并巩固与Lambda rex基因（赋予rex基因）相关的表型以及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在阐明Lambda T4rII排斥机制方面的初步成功。这里概述的研究不仅有助于提高我们对经典遗传奥秘的理解，并提供有关这个神秘系统的重要科学信息，但更重要的是，翻译这些发现-识别类似的人类病毒编码排斥系统，可能用于开发新的抗病毒药物。该计划还为培养具有分子遗传学、噬菌体生物学、核酸和蛋白质生物化学综合专业知识的高素质人才提供了坚实的科学平台。

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.