Publication Date

Spring 4-27-2021

School

School of Health Sciences

Major

Biology: Biomedical Sciences

Keywords

recombinant protein purification, antibody, co-immunoprecipitation, western blot, folate, gene expression, silver stain

Disciplines

Biochemistry | Cell Biology | Molecular Biology | Molecular, Genetic, and Biochemical Nutrition | Molecular Genetics

Abstract

A previous study that introduced dietary folate to mice in the form of folic acid to determine if gene activity would be altered based on this biological molecule demonstrated that mice without folic acid had cognition deficits, and this phenomenon was correlated with altered gene expression in their brains. The included bioinformatic analysis revealed two main transcription factors that bind to proteins in the nucleus, and one is known as the Zinc Finger Protein 410 (Zfp410). Due to the lack of literature explaining the function of this transcription factor, this project is intended to analyze Zfp410 in detail from scratch. Zfp410 cDNA sequence was purchased along with a plasmid vector, transferred to E. coli, and used to purify the recombinant protein. The purified protein was then injected into a rabbit for antibody production, and the collected Zfp410 antibody was characterized using several molecular techniques. Within high efficiency of targeting its antigen, Zfp410 antibody was used to identify any interacting proteins of Zfp410 by co-immunoprecipitation along with silver stain and western blot. The results demonstrated several Zfp410 interacting partners which suggested transcription factor Zfp410’s role in folate-dependent genes’ regulations.

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