- This event has passed.
Seminar: Emily Brooks: Bioinformatic-assisted Determination of Novel Constitutive Soybean Cis-Regulatory Elements
August 19, 2022 | 10:00 am - 11:00 am
Bioinformatic-assisted Determination of Novel Constitutive Soybean Cis-Regulatory Elements
Emily Brooks, MS Seminar
Under the direction of Dr. Wusheng Liu
Friday, August 19, 2022 at 10:00 am
Join Zoom Meeting https://ncsu.zoom.us/j/99241760094?pwd=Zm1taFkrQ3lCMGtPWDRUdTV3R29CUT09
Meeting ID: 992 4176 0094
Passcode: 917233
Summary:
Determining the promoter motifs involved in regulating transcription, the first process of gene expression, is critical for synthetic promoter engineering. A gene’s promoter contains cis-regulatory elements, or motifs, which are transcription factor (TF) binding sites that bind TFs to initiate and drive transcription. Many bioinformatic tools have been developed for determining statistically overrepresented regions that are shared across groups of promoters. Combinations of multiple bioinformatic tools can lead to improved accuracy in determining motifs conferring biological activity. In the present study, we compiled a set of 11 known soybean constitutive gene promoters under the assumption that some of the promoters are regulated by the same transcription factor(s). Seven bioinformatic tools capable of de novo motif discovery were used to determine potentially shared motifs within the promoters, which were then mapped back to the original promoter sequences. A total of 64 overlapping motif regions (OMRs) were commonly detected amongst the 11 constitutive promoters, and each OMR was cloned individually in front of a minimal 35S promoter driving the GUSPlus reporter gene expression. Tobacco leaf agroinfiltration assay and subsequent quantitative GUS activity assay was used to determine each OMR’s ability to drive gene expression. We found 20 out of the 64 OMRs drove significantly higher gene expression than that conferred only by the minimal 35S promoter, and 11 of the 20 OMRs drove expression at a level two times or greater than that by the minimal 35S promoter. These functional OMRs are strong candidates for further characterization using stable transgenics, identification of the actual core motif sequences within the OMRs, and synthetic constitutive promoter engineering.