Abstract Summary/Description
The global rise of Neisseria gonorrhoeae necessitates a deeper exploration of its biology to address emerging challenges in combating gonorrhea. Optimizing complementation experiments is crucial for understanding the bacterium's complex biology and unraveling genetic factors related to challenges, particularly focusing the requirements for optimum protein translation initiation. Our study investigates the influence of different spacer sequence lengths between the ribosome binding site and start codon on super folder Green Fluorescent Protein (sfGFP) production, serving as a visual readout for protein expression. Colony screening reveals that increasing spacer lengths from 5 to 8 nucleotides leads to a gradual elevation in sfGFP production, while spacers exceeding 8 nucleotides result in decreased production of sfGFP. Ongoing experiments, including fluorescence intensity measurement, flow cytometry, and Western blot techniques, will enable a comprehensive evaluation of protein production across different spacer variants. These findings contribute to a deeper understanding of N. gonorrhoeae molecular biology and offer practical insights for refining complementation and protein production experiments in future studies.
