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。尽管在这一领域的工作已经使人们对基因调控、DNA复制和重组有了突破性的了解，但仍有许多事情需要了解，许多具有潜在开发价值的谜团仍未解开。其中一个特别有意义的是Lambda T4rII排除(Rex)表型。这种表型是一种复杂的神秘现象，定义为噬菌体T4快速裂解突变体(称为RII突变体)无法在携带整合到大肠杆菌染色体中的休眠Lambda噬菌体的大肠杆菌细胞草坪上形成斑块。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，也是迄今为止定位最彻底的基因，但RII逃避Rex的机制和RII排斥表型本身仍然不清楚。 这种表型的阐明可能会将我们引向类似的真核病毒排除系统，这些系统可能被用于开发新的抗病毒策略。这项建议旨在确保关键人员和设备的安全，以执行这些遗传和生化分析，寻求显著扩展我们的理解，并巩固与Lambda rex基因(赋予Rex)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在阐明Lambda T4rII排斥机制方面取得了初步成功。这里概述的研究不仅有助于增进我们对经典遗传之谜的理解，并提供关于这一神秘系统的重要科学信息，更重要的是，将这些发现转化为识别类似的人类病毒编码的排斥系统，这些系统可能被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、噬菌体生物学以及核酸和蛋白质生物化学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.