Presentation Type
Poster Presentation
Category
STEM (science, technology, engineering, mathematics)
Abstract/Artist Statement
Investigating proteins involved in driving cancer progression is crucial for advancing therapeutic strategies. PRDM3 (PR/SET domain-containing 3) has emerged as a key player in sustaining ovarian tumors. To facilitate an in-depth study of PRDM3, obtaining the protein in substantial quantities with a high-level purity through recombinant expression is imperative.
The methodology comprises a series of steps: integrating the PRDM3 gene fused with an affinity tag into a plasmid; transferring the engineered plasmid into bacterial cells; expressing the affinity-tagged protein (fusion protein) within the bacterial cells and harvesting them. Cell lysis and protein extraction are then conducted, followed by affinity chromatography for fusion protein purification and subsequent cleavage of the affinity tag. Gel filtration chromatography is used for additional purification, resulting in a homogeneous PRDM3 protein for detailed structural analyses.
Achieving maximum purity and a high yield in the purification of PRDM3 poses challenges in protein extraction, tag cleavage and homogeneity attainment. This can significantly impact the feasibility of subsequent protein studies, necessitating optimization strategies.
This study aims to address these challenges by refining the protein purification methods for PRDM3, facilitating a more precise understanding of its interactions with other proteins and its role in oncogenic processes. Standard chromatographic separations, such as affinity and gel filtration chromatography, combined with an affinity tag and the use of detergent, have been utilized to optimize protein extraction. Simultaneously, changes in cleaving conditions have been introduced to minimize protein precipitation post-tag cleavage. The introduction of these combined enhancements distinguishes this study from conventional methods and demonstrates a novel strategy for optimizing PRDM3 protein extraction and purification, ultimately ensuring its isolation with optimal purity and yield.
In conclusion, this study lays the groundwork for more in-depth investigations into the molecular interplay between PRDM3 and other proteins- potentially leading to advances in therapeutics for ovarian cancer.
Mentor Name
Klara Briknarova
Personal Statement
This research holds immense scientific significance as it delves into a critical aspect of cancer biology, unraveling the role of PRDM3 in ovarian tumors. Its impact extends to clinical prognosis, notably with the identification of PRDM3 protein’s interactions linked to unfavorable outcomes in gynecological cancers, thereby elevating its relevance to patient care. By investigating the molecular mechanism of PRDM3, this study aims to provide invaluable insights that could reshape the landscape of cancer research and guide therapeutic strategies. The innovative methodology employed benefits the scientific community and contributes to the global fight against cancer by enhancing diagnostics and treatments. The rigorous approach, including a detailed optimization of protein purification, not only enhances credibility but also showcases technical innovation. This work transcends the realm of science, promising real-world impact on human health and well-being, highlighting its translational value. The methodological approach involves utilizing recombinant expression to generate a substantial quantity of PRDM3, laying the foundation for comprehensive structural and functional analyses. Recognizing the prognostic relevance of PRDM3 protein interactions in gynecological cancers, the research aims to unravel the underlying molecular mechanisms and understand their implications for disease outcomes. A noteworthy aspect of this study is the candid acknowledgment of challenges encountered in achieving high purity and yield during the purification process. The dedicated effort and time to refine and optimize purification methods emphasizes the commitment to surmounting these obstacles. This optimization is not only pivotal for the success of the present study but also holds implications for structural analysis and functional assays. The goal of the study extends beyond obtaining PRDM3 with optimal purity; it aims to pave the way for more profound investigations into the molecular mechanisms underpinning cancer progression, thereby contributing to advancements in our understanding of these complex diseases.
Video presentation
PRDM3 Protein Purification: An Optimized Approach for Structural Understanding
UC North Ballroom
Investigating proteins involved in driving cancer progression is crucial for advancing therapeutic strategies. PRDM3 (PR/SET domain-containing 3) has emerged as a key player in sustaining ovarian tumors. To facilitate an in-depth study of PRDM3, obtaining the protein in substantial quantities with a high-level purity through recombinant expression is imperative.
The methodology comprises a series of steps: integrating the PRDM3 gene fused with an affinity tag into a plasmid; transferring the engineered plasmid into bacterial cells; expressing the affinity-tagged protein (fusion protein) within the bacterial cells and harvesting them. Cell lysis and protein extraction are then conducted, followed by affinity chromatography for fusion protein purification and subsequent cleavage of the affinity tag. Gel filtration chromatography is used for additional purification, resulting in a homogeneous PRDM3 protein for detailed structural analyses.
Achieving maximum purity and a high yield in the purification of PRDM3 poses challenges in protein extraction, tag cleavage and homogeneity attainment. This can significantly impact the feasibility of subsequent protein studies, necessitating optimization strategies.
This study aims to address these challenges by refining the protein purification methods for PRDM3, facilitating a more precise understanding of its interactions with other proteins and its role in oncogenic processes. Standard chromatographic separations, such as affinity and gel filtration chromatography, combined with an affinity tag and the use of detergent, have been utilized to optimize protein extraction. Simultaneously, changes in cleaving conditions have been introduced to minimize protein precipitation post-tag cleavage. The introduction of these combined enhancements distinguishes this study from conventional methods and demonstrates a novel strategy for optimizing PRDM3 protein extraction and purification, ultimately ensuring its isolation with optimal purity and yield.
In conclusion, this study lays the groundwork for more in-depth investigations into the molecular interplay between PRDM3 and other proteins- potentially leading to advances in therapeutics for ovarian cancer.