Investigating the Expression and Purification of Protein TTHA0508 in the Foundational Organism Thermus thermophilus HB8
Disciplines
Biochemistry, Biophysics, and Structural Biology
Abstract (300 words maximum)
Investigating the Expression and Purification of Protein TTHA0508 in Foundational Organism Thermus thermophilus HB8.
Foundational organisms are used in research to provide fundamental concepts or processes for further investigation. Thermus thermophilus is a foundational thermophile, an organism whose ideal temperature conditions are higher than other organisms. This investigation specifically studies Thermus thermophilus HB8 and the protein expression of TTHA0508 in E. coli Rosetta 2 strain (DE3) pLysS through bacterial transformations. The protein of interest, TTHA0508, is a transcription factor; a protein that contributes to the process of transcription and the overall gene expression of that organism. Primarily, this research analyzes the ideal conditions for TTHA0508 protein expression. Following the expression of the protein 0508, our transformant is used to find optimal conditions for TTHA0508 purification. The goal of investigating the expression and purification of TTHA0508 protein is to provide understanding for later research in Restriction Endonuclease Protection, Selection, and Amplification (REPSA) of Thermus thermophilus HB8. The protein TTHA0508 is symmetrical and contains two MeR family proteins, suggesting multiple unique gene interactions, unlike many transcription factors where the protein contains only one MeR family protein. REPSA of T. thermophilus provides information about what gene(s) TTHA0508 interacts with. The ideal protein expression procedure follows a bacterial transformation of the E. coli cells with the plasmid containing the TTHA0508 gene and inducing the cells using IPTG, a metabolite that inhibits a repressor protein on the LAC operon and thus induces expression. Continuing with the induced cells, purification entails suspending the transformed E. coli cells in a high sodium chloride salt solution and then a series of centrifugation and heat treatments. The results are analyzed through a series of protein gel electrophoresis and provide foundational information to further investigate the TTHA0508 transcription factor through REPSA.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Michael Van Dyke
Investigating the Expression and Purification of Protein TTHA0508 in the Foundational Organism Thermus thermophilus HB8
Investigating the Expression and Purification of Protein TTHA0508 in Foundational Organism Thermus thermophilus HB8.
Foundational organisms are used in research to provide fundamental concepts or processes for further investigation. Thermus thermophilus is a foundational thermophile, an organism whose ideal temperature conditions are higher than other organisms. This investigation specifically studies Thermus thermophilus HB8 and the protein expression of TTHA0508 in E. coli Rosetta 2 strain (DE3) pLysS through bacterial transformations. The protein of interest, TTHA0508, is a transcription factor; a protein that contributes to the process of transcription and the overall gene expression of that organism. Primarily, this research analyzes the ideal conditions for TTHA0508 protein expression. Following the expression of the protein 0508, our transformant is used to find optimal conditions for TTHA0508 purification. The goal of investigating the expression and purification of TTHA0508 protein is to provide understanding for later research in Restriction Endonuclease Protection, Selection, and Amplification (REPSA) of Thermus thermophilus HB8. The protein TTHA0508 is symmetrical and contains two MeR family proteins, suggesting multiple unique gene interactions, unlike many transcription factors where the protein contains only one MeR family protein. REPSA of T. thermophilus provides information about what gene(s) TTHA0508 interacts with. The ideal protein expression procedure follows a bacterial transformation of the E. coli cells with the plasmid containing the TTHA0508 gene and inducing the cells using IPTG, a metabolite that inhibits a repressor protein on the LAC operon and thus induces expression. Continuing with the induced cells, purification entails suspending the transformed E. coli cells in a high sodium chloride salt solution and then a series of centrifugation and heat treatments. The results are analyzed through a series of protein gel electrophoresis and provide foundational information to further investigate the TTHA0508 transcription factor through REPSA.