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。

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.