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Presenter: Hayley Mello

Faculty Sponsor: Jason M. Brown

School: Salem State University

Research Area: Biology

Session: Poster Session 3, 1:15 PM - 2:00 PM, Auditorium, A34

ABSTRACT

Following the loss of cilia, Chlamydomonas reinhardtii can regrow them by inducing hundreds of genes. This research aims to investigate the process of gene regulation as cilia regrow. Luminescent reporter genes, such as luciferase, allow for monitoring the activation of a selected gene’s promoter via light emission. Quantitative PCR (qPCR) enables a more direct measurement of endogenous gene expression. We are working with a reporter strain where the promoter from LC8, a gene known to be upregulated during cilia regeneration, is connected to the Gaussia luciferase coding region. Cells were pH-shocked from pH 7 to pH 4, then back to pH 7, to trigger deciliation and allow for regrowth. Samples were frozen at 0, 30, and 60 minutes for analysis by luminescence assay and isolation of RNA for qPCR. Separate samples at each time point were fixed with iodine and imaged via phase contrast microscopy for cilia measurement. The results of the assays and qPCR will be compared to evaluate the expression of the target gene. Analysis of luminescence assay data indicates activation of the LC8 promoter in wild-type cells post-pH shock. Preliminary evaluation of qPCR values suggests expected amplification of the target gene. Further analysis will be conducted to determine what can be concluded about the gene regulation observed in C. reinhardtii wild-type and mutant cells.