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Title

Μελέτη της κινητικότητας του μεταθετού στοιχείου Μinos σε βακτήρια

Author

Κυριακάκης, Εμμανουήλ

Thesis advisor

Σαββάκης, Χαράλαμπος

Abstract

The transposable element Minos, belongs to the Tc1/mariner superfamily of transposable elements and was originally identified in Drosophila hydei. Several members of this superfamily have been found to be active, even in hosts evolutionarily diverse from the elements original host. The presence of an active transposase, the enzyme that catalyzes the transposition, is the only requirement for transposition to occur. This characteristic makes elements like Minos a valuable genetic tool. As shown in the present study, the Minos transposable element is not able to transpose inside E.coli under the conditions used, or the efficiency of transposition is so low, that it can not be detected. With the excision assay and the transposition assays which were used in this study. That was probably due to many different factors, such as the potency of the promoter that drives the expression of the transposase and/or the stability of the transcript of the transposase orthe stability of the protein itself in E.coli. Finally, with the use of error prone PCR it was feasible to introduce mutations into the transposase by random mutagenesis. Two libraries were created and were screened for active variants of the transposase using a papillation assay to trace transposition events. The preliminary experiments have not shown any apparent activity.

Language

Greek

Subject

Genetics

Γενετική

Issue date

2005-12-12

Date available

2006-11-20

Collection

School/Department--School of Medicine--Department of Medicine--Post-graduate theses

Type of Work--Post-graduate theses