In rare cases, bacterial transcripts begin with the start codon and thus do not contain a 5’ untranslated sequence. Because they lack this untranslated leading sequence, such unorthodox transcripts are considered “leaderless”. How translation initiates under these circumstances has not been well characterized. In this project, we investigate factors influencing the expression of leaderless mRNA (lmRNA) in Agrobacterium. We chose this type of bacteria because data from other studies suggests that alphaproteobacteria, such as Agrobacterium, often use leaderless translation more frequently than E. coli. Using a leaderless reporter system, we will compare the levels of expression of lmRNA in E. coli and Agrobacterium. Following this comparison, we will perform a whole genome mutagenesis in Agrobacterium and subsequent screening of the transformed mutants. By carrying out whole genome sequencing of mutants with varying levels of expression of lmRNA, we expect to identify various ribosomal mutations that affect leaderless translation in Agrobacterium. These findings will contribute to our fragmented understanding of the mechanism behind leaderless translation in bacteria.
Chromoproteins
We are working on a project to mutagenize chromoprotein plasmids, aiming to alter their color expression or improve their maturation time and intensity. Chromoproteins are proteins that produce vibrant, visible colors without the need for enzymatic reactions or cofactors. By introducing targeted mutations into the chromoprotein-encoding genes, we seek to identify variants with enhanced or novel properties. We are specifically working with chromoproteins that produce hot pink, bright yellow, deep yellow, and blue colors and have seen that our targeted mutagenesis does produce colors different from the original protein. We hope to add novel colors or faster maturing proteins to the agar art scene.
