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Yeast Transactivation Assay Lab Report

1003 Words5 Pages

This purpose of this experiment was to determine if a new yeast transactivation assay was able to determine estrogenic activity in estrogenic compounds. Throughout this experiment, researchers compared the detection ability of the new transactivation assay with the conventional transactivation assay. The new transactivation assay plasmid replaced the lacZ gene with GFP (green fluorescent protein) gene, and by doing so simplifies the process of determining estrogenic activity. The advantages of using GFP as the reporter instead of beta-galactosidase, which is the protein that the lacZ gene codes for, are numerous. The presence of beta-galactosidase is detected by photometric measurement, which requires the yeast cells to be disrupted which is both a time-consuming error and also a source of error. GFP is stable in a large pH range and is highly soluble, making it adaptable to a wide …show more content…

When 17beta-estradiol was added to the yeast cells, most emitted green fluorescence. There seemed to be a direct relationship with the magnitude of GFP expression and the concentration of 17beta-estradiol concentration. The results show that does GFP function as a satisfactory reporter gene for estrogen in this transactivation assay, satisfactory enough to replace the conventional beta-galactosidase transactivation assay. This study proves that GFP can replace beta-galactosidase as a reporter plasmid- a beneficial follow up study would be to see if there was a way to measure the level of enzyme activity in estrogenic compounds without transforming other genes upstream from the gene of interest, which in this case would be estrogen receptor alpha. If scientists can investigate ways to simply measure the activity of that expression plasmid, it would simplify this entire process and diminish the need for transactivation assays

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