Investigating the Rex exclusion genes of bacteriophage Lambda

点击翻译按钮获取中文摘要

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.