Protocol to evaluate translation inhibition in Candida species using fluorescence microscopy and flow cytometry

Abstract

Summary: Translation inhibitors have therapeutic potential against Candida species. Here, we present a protocol to measure translation inhibition in Candida spp. We describe steps for employing an alkynylated methionine analog, L-homopropargylglycine (HPG), that becomes incorporated into newly synthesized proteins. We then detail procedures to perform a click reaction of the alkyne with a fluorescent azide, which is visualized using fluorescence microscopy and quantified by flow cytometry.For complete details on the use and execution of this protocol, please refer to Puumala et al.,1 Fu et al.,2 and Iyer et al.3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• genetics
• microbiology
• microscopy