Croucher, Kristina*, and Michael Foulk
Mercyhurst University, Erie, PA 16546.
Identification and characterization of the Sciara coprophila cold tolerance gene, Frost (ScFst).-
Organisms in the wild routinely encounter extreme environmental conditions. Seasonal cold temperatures pose a threat to insects in particular; thus, they have developed mechanisms to recover from cold shock. Recently, the effects of cold on the fruit fly
Drosophila melanogaster have been studied and a number of genes responsible for cold tolerance have been identified. One of these genes is
Frost (
Fst). While the specific function of
Frost has yet to be determined, expression of the gene has been shown to rise during the period of recovery after exposure to cold temperatures. In this study, we have identified and begun to characterize a potential
Frost ortholog in the fungus gnat,
Sciara coprophila. The
D. melanogaster Frost sequence was collected from NCBI and BLAST was used to identify a potential
Frost ortholog in the
S. coprophila genome. Several candidates were initially identified and a reciprocal BLAST search with each of the candidates was completed to identify a potential
S. coprophila Frost ortholog, which was named the putative
S. coprophila Frost (ScFst) gene. Total RNA was extracted from adult female flies subjected to a three-hour cold stress then allowed to recover for 0, 1, 2, and 3 hours. Quantitative Real-Time Polymerase Chain Reaction (qPCR) was used to analyze putative
ScFst expression at each of these time points. Expression increased for at least the first hour time point. Future plans are to analyze
ScFst expression during recovery from cold stress in other developmental stages, such as embryo, larvae, and pupae. We are also in the process of cloning the putative
ScFst mRNA into a topocloning vector known as pCR2.1 and then subclone the sequence in frame with GFP. (
31)
Poster Link