Presenter Information

Benjamin V. HickeyFollow

Presentation Type

Poster

Faculty Mentor’s Full Name

Ekaterina Voronina

Faculty Mentor’s Department

CBSD

Abstract

FBF-1 and FBF-2 are proteins within the PUF protein family that are required for stem cell maintenance in Caenorhabditis elegans. FBF-1 and FBF-2 exhibit mRNA binding activity and are involved in localization, activation and repression of their target mRNA’s. The two are similar in sequence with the exception of four Variable Regions (VR’s). FBF-2 localizes to P granules in germ cells of the C. elegans while FBF-1 does not. We propose that the different localization patterns exhibited between the FBF-1 and 2 are due to these VR’s. Analysis of which VR or combination of VR’s is responsible for this difference in localization was undertaken through chimeric protein assembly and insertion into the C. elegans genome. Assembly of DNA encoding chimeric proteins with various VR’s and a tagging Fluorescent Protein (GFP) present was achieved through fusion Polymerase Chain Reaction (PCR) and BP/LR clonase plasmid assembly. Introduction of chimeric DNA constructs is in progress, and done through Crispr-CAS-9 genome editing. Expression of the modified proteins and assessment of localization patterns will be carried out using GFP visualization. The poster will discuss our observations and preliminary conclusions.

Category

Life Sciences

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Apr 27th, 11:00 AM Apr 27th, 12:00 PM

Using Chimeric Proteins to Determine Basis of FBF-2 Localization

UC South Ballroom

FBF-1 and FBF-2 are proteins within the PUF protein family that are required for stem cell maintenance in Caenorhabditis elegans. FBF-1 and FBF-2 exhibit mRNA binding activity and are involved in localization, activation and repression of their target mRNA’s. The two are similar in sequence with the exception of four Variable Regions (VR’s). FBF-2 localizes to P granules in germ cells of the C. elegans while FBF-1 does not. We propose that the different localization patterns exhibited between the FBF-1 and 2 are due to these VR’s. Analysis of which VR or combination of VR’s is responsible for this difference in localization was undertaken through chimeric protein assembly and insertion into the C. elegans genome. Assembly of DNA encoding chimeric proteins with various VR’s and a tagging Fluorescent Protein (GFP) present was achieved through fusion Polymerase Chain Reaction (PCR) and BP/LR clonase plasmid assembly. Introduction of chimeric DNA constructs is in progress, and done through Crispr-CAS-9 genome editing. Expression of the modified proteins and assessment of localization patterns will be carried out using GFP visualization. The poster will discuss our observations and preliminary conclusions.