Identifying protein-protein interactions of a cell cycle regulator
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The role of anachronism (ana) protein in stem cell division of Drosophila melanogaster was examined. Synthesis of identifiable ana protein was necessary. The identifying method exploited was that of antibody tagging using a myc epitope or a poly-Histidine region. Analysis of previously utilized complementary DNA (cDNA) sequences showed errors in the alignment of the reading frame. Methods were employed to clone the required ana cDNA sequence into an expression vector (pBAD/Myc-HisA) within the correct reading frame. Restriction endonucleases were then used to confirm that the cDNA sequence and the expression vector had ligated properly. Analysis of the results from the single- and double-restriction digests showed that the ana cDNA (1.6 kilobases) was successfully cloned and ligated into the pBAD/Myc-HisA (4.1 kilobases) expression vector. The ana cDNA was also successfully cloned in frame with the sequence coding for the myc epitope and the poly-Histidine region. This data provides groundwork for the expression of identifiable ana protein.
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Includes bibliographical references (leaves 27-28).
Amos, Joseph Edward (2001). Identifying protein-protein interactions of a cell cycle regulator. Texas A&M University. Available electronically from