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Purification of Mitotic Phosphatase Cdc14p

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dc.contributor.author Torres, Emily
dc.contributor.author Simons, Kim
dc.date.accessioned 2011-10-20T15:30:54Z
dc.date.available 2011-10-20T15:30:54Z
dc.date.created June 2010 en_US
dc.date.issued 2011-10-20
dc.identifier.uri http://hdl.handle.net/123456789/117
dc.description.abstract Two methods were employed to achieve soluble expression of Cdc14p. The first being the retrieval of CDC14 from a pET28 expression plasmid and insertion into a GST plasmid by use of the restriction enzyme NdeI. Before this could occur, alteration of an internal NdeI recognition sequence was attempted via site-directed mutagenesis. The second method being a process of exposing cells containing pET28 to a solution 10% sarkosyl or of 2% sarkosyl 2% triton X-100 and 20 mM CHAPS to disrupt any possible inclusion bodies of properly folded Cdc14p. Once soluble protein is extracted by either method, two assays will be preformed to ensure enzymatic activity and specificity will be examined. en_US
dc.language.iso en_US en_US
dc.subject CDC14 en_US
dc.subject molecular cloning en_US
dc.subject phosphatase en_US
dc.subject cell cycle en_US
dc.title Purification of Mitotic Phosphatase Cdc14p en_US
dc.type Technical Report en_US
dc.college las en_US
dc.academic.area Biochemistry en_US
dc.advisor Simons, Kim en_US
dc.department physical sciences en_US

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