Isolation and Amplification of the strK gene of Streptomyces griseus
Evers, Danielle. “Isolation and Amplification of the strK gene of Streptomyces griseus”, Boston College, 2004. http://hdl.handle.net/2345/480.
My project focused on the study of streptomycin-6-phosphatase, an enzyme that has shown specificity for the substrate streptomycin-6-phosphate. The study included attempts at isolation, purification, and amplification of strK, the gene encoding this phosphatase. These efforts were made in order to come closer to gaining an understanding of the specificity of this enzyme especially in comparison to alkaline phosphatase, a well-documented unspecific phosphate with notable similarity of sequence and structure. The major question which developed into the study undertaken this year was: “How does a specific phosphatase compare to the unspecific alkaline phosphatase (AP)?” This is a longterm project that could take years to come close to elucidating an answer to. My approach to this large question, under direction of Dr. Evan Kantrowitz, was to begin with streptomycin-6-phosphatase, the product of the strK gene in Streptomyces, and embark on a study that would lead to greater understanding of this specific phosphatase. This undertaking was pursued based upon previous publications identifying conservation of amino acid sequence between the two phosphatases. This similarity was most poignantly found at sites found in AP noted as contained in the active site and in taking part in metal binding. From this information, the question for my study became, “How much can I learn about strK gene product during my time of study so that progress is made toward shedding light on the complex question posed above?”.