Understanding How bus-1 is Regulated at the Molecular Level

Abstract

The nematode species Caenorhabditis elegans is especially suitable for studying a range of genetic and biological questions. Here, we have used C. elegans to study host/pathogen interactions. Screens of the C. elegans genome have revealed bus-1 as a gene whose product affects the nematode’s sensitivity to the bacterial pathogen Microbacterium nematophilum (Gravato-Nobre and Hodgkin, 2006). Mutations in bus-1 prevent M. nematophilum attachment and subsequent post-anal swelling in C. elegans. We aim to learn how bus-1 is regulated at the molecular level by transcription factor EGL-38 to influence expression of phenotypes in the anal region. In 2013, a colleague Benjamin Kaumeyer made a bus-1 reporter construct to determine its expression pattern. This was done by cloning a 1500 base pair fragment of DNA located 5’ to the start of the bus-1 gene into a GFP reporter. We have conducted a deletion analysis of Kaumeyer’s construct by removing four DNA fragments of various lengths upstream of the bus-1 gene to learn what region of the bus-1 promoter is regulated by EGL-38. The deletion clones were purified and injected into worms to reveal a 274 nucleotide potential EGL-38 binding site on bus-1.