Κατασκευή δικιστρονικών και μονοκιστρονικών φορέων έκφρασης θηλαστικών, συμβατών με το σύστημα μετάθεσης “sleeping beauty” (Bachelor thesis)

Ρουμελιώτης, Σπυρίδων

In the present thesis, the construction of a set of composite mammalian expression vectors is described. The designed vectors can be used to stable transfect cells with the “sleeping beauty” transposition system. In order to accomplish that, all of them contain two ITR (Inverted Terminal Repeats) sequences recognised by the transposase of the sleeping beauty transposon. This enzyme transposes the DNA region included between these two sequences, into the cellular genome. Furthermore, these vectors can provide high expression levels of one or two genes of interest. We have created two alternate versions (Figure 1); the first employs a ubiquitous promoter in order to achieve constitutive, high expression while the second uses a neurospecific promoter which drives high expression levels in neural cells. Moreover, each one of the two versions is available in three variations which differ in the structure of the transcriptional unit used. The first variation can be used for over-expression of one gene of interest. The second contains the EMCVIRES and thus, can be employed for the simultaneous expression of a gene of interest and a reporter gene (either eGFP, or mCherry). The third one (also contains the EMCV-IRES) can be utilised for simultaneous expression of two different genes of interest.
Institution and School/Department of submitter: Δημοκρίτειο Πανεπιστήμιο Θράκης. Σχολή Επιστημών Υγείας. Τμήμα Μοριακής Βιολογίας και Γενετικής
Subject classification: Cells
Keywords: Bicistronic,Sleeping beauty,Transfectance,Δικιστρονικοί φορείς έκφρασης,Σύστημα μετάθεσης,Σταθερή διαμόλυνση
URI: https://repo.lib.duth.gr/jspui/handle/123456789/17195
http://dx.doi.org/10.26257/heal.duth.15929
Appears in Collections:ΤΜΗΜΑ ΜΟΡΙΑΚΗΣ ΒΙΟΛΟΓΙΑΣ & ΓΕΝΕΤΙΚΗΣ

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http://dx.doi.org/10.26257/heal.duth.15929
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