dc.contributor.author	Col, Bekir	en
dc.contributor.committeechair	Rutherford, Charles L.	en
dc.contributor.committeemember	Esen, Asim	en
dc.contributor.committeemember	Stevens, Ann M.	en
dc.contributor.committeemember	Favis, Reyna	en
dc.contributor.department	Biology (Molecular and Cellular)	en
dc.date.accessioned	2014-03-14T20:39:09Z	en
dc.date.adate	1998-01-07	en
dc.date.available	2014-03-14T20:39:09Z	en
dc.date.issued	1997-12-12	en
dc.date.rdate	1999-01-07	en
dc.date.sdate	1997-12-12	en
dc.description.abstract	Glycogen phosphorylase 2 (gp-2) is a key enzyme during the development of Dictyostelium discoideum. The gp-2 enzyme breaks down glycogen into glucose monomers that are subsequently used to synthesize the terminal end products of cellular differentiation. This gene is an ideal candidate for studying the process of selective gene expression because its product figures so prominently in the development of this organism, implying a dependable control mechanism responsible for its developmentally regulated expression. I present in this thesis the identification of several putative cis-acting elements of gp-2 as revealed through footprint analysis. Due to the extreme AT-bias characteristic of Dictyostelium promoters, footprinting conditions required intensive optimization with respect to template, nonspecific competitor, source of protein extract and DNase I digestion. Using an endlabeled fragment containing seven repeated sequences (3 TA boxes [TAATTATA], 2 TAG boxes [TAAAAATGGT] and 2 C boxes [ACCCACT]), purified replication protein A and several developmental nuclear extracts were tested for DNA binding activity. Small footprints were observed on the TAG and C boxes of the promoter for both protein sources. However, using a more sensitive footprinting strategy involving multiple rounds of primer extension, larger footprints spanning the same promoter regions were detected. In both cases, the appearance of the footprints coincided with the documented transcriptional activity of the gene. It can be concluded from the data obtained that the TAG and C boxes are very likely cis-acting elements involved in the regulation of gp-2 expression.	en
dc.description.degree	Master of Science	en
dc.identifier.other	etd-0598-12052	en
dc.identifier.sourceurl	http://scholar.lib.vt.edu/theses/available/etd-0598-12052/	en
dc.identifier.uri	http://hdl.handle.net/10919/33366	en
dc.publisher	Virginia Tech	en
dc.relation.haspart	title_abstract.pdf	en
dc.relation.haspart	sections.pdf	en
dc.relation.haspart	table_A_B.pdf	en
dc.relation.haspart	figure_4.1_and_4.2.pdf	en
dc.relation.haspart	figure_3.1___3.16.pdf	en
dc.relation.haspart	figure_1.3.pdf	en
dc.relation.haspart	figure_2.2.pdf	en
dc.relation.haspart	Figure_1.2.pdf	en
dc.relation.haspart	figure_1.1.pdf	en
dc.relation.haspart	acknowledgement_table_of_contents.pdf	en
dc.relation.haspart	figure1.4.pdf	en
dc.relation.haspart	figure2.1.pdf	en
dc.rights	In Copyright	en
dc.rights.uri	http://rightsstatements.org/vocab/InC/1.0/	en
dc.subject	cell differentiation	en
dc.subject	Dictyostelium discoideum	en
dc.subject	DNase I footprint analysis	en
dc.subject	transcription	en
dc.subject	gene expression	en
dc.subject	AT-rich	en
dc.subject	looping	en
dc.subject	large footprints	en
dc.subject	replication protein A	en
dc.title	Footprint Analysis of the Transcriptional Control of Glycogen Phosphorylase 2 in Dictyostelium Discoideum	en
dc.type	Thesis	en
thesis.degree.discipline	Biology (Molecular and Cellular)	en
thesis.degree.grantor	Virginia Polytechnic Institute and State University	en
thesis.degree.level	masters	en
thesis.degree.name	Master of Science	en

Footprint Analysis of the Transcriptional Control of Glycogen Phosphorylase 2 in Dictyostelium Discoideum

Files

Original bundle

Collections