TRANSFORMATION PROTOCOLS IN CHLAMYDOMONAS REINHARDTII FOR LABORATORY INSTRUCTION
Corbett, Jonathan (Biology)
Faculty Mentors: Ross Koning & Mike Adams
Chlamydomonas reinhardtii is a green alga commonly used as a model organism in genetics and cellular biology research and teaching laboratories. Its benefits include the ability to grow mixotrophically on inexpensive media and the availability of genome sequences and markers. Several plasmids, optimized for Chlamydomonas, have been synthesized containing reporter genes encoding for bioluminescence. This work employed protocols for introducing bioluminescence genes from the jellyfish Aequorea victoria and the copepod Gaussia princeps using equipment commonly found in laboratories at secondary schools and smaller universities. GFP expression, easily visible in E. coli under UV light, was confirmed in Chlamydomonas only by confocal microscopy. Visualization with the naked eye was presumably masked by quenching due to chlorophyll a. Bioluminescence from the Gaussia luciferase reaction was detected in a microtiter plate reader, but was similarly invisible to the unaided eye. Additionally, transformation frequencies from all protocols were not consistent enough for effective use in an educational laboratory. Because all of these approaches required expensive technologies, none proved capable of demonstrating transformation in a typical instructional laboratory environment.