Unit:
Τομέας Βιοχημείας Μοριακής ΒιολογίαςLibrary of the School of Science
Author:
Τσατσαρούνος Σωτήρης
Dissertation committee:
Λεκανίδου Ρένα Ομότιμη Καθηγήτρια ΕΚΠΑ (επιβλέπουσα), Ροδάκης Γεώργιος Καθηγητής ΕΚΠΑ, Swevers Luc Ερευνητής Α’, ΕΚΕΦΕ «Δημόκριτος»
Original Title:
Μοριακή ανάλυση της διαφορικής έκφρασης γονιδίων σε λεπιδόπτερα έντομα: Μελέτη της "αρχιτεκτονικής" του υποκινητή του γονιδιακού ζεύγους Α/Β.L9 στο μεταξοσκώληκα bombyx mori μέσω ηλεκτροδιάτρησης
Translated title:
Molecular analysis of differential gene expression in Lepidoptera insects:“Architecture” analysis of the A/B.L9 gene pair promoter in the silkmoth Bombyx mori via electroporation
Summary:
A useful system for studying the molecular mechanisms, implicated in
differential gene expression, is choriogenesis (eggshell formation) in Bombyx
mori silkmoth.
The primary aim of this thesis was to investigate how promoter “architecture”
(number, orientation and position of transcriptional factor -C/EBP, GATA, HMGA,
CHD1- binding sites) facilitates developmentally accurate chorion gene
regulation. Towards this purpose, several mutated (and non-mutated) promoter
regions of different developmental specificity chorion genes, were cloned
upstream of a reporter gene (lacZ) to introduce these plasmid-constructs into
silkmoth follicle epithelial cells via electroporation as an efficient and
quick method for transient expression (on ex vivo developing follicles).
At first, electroporation parameters were optimized (250Volts/cm current,
25msec and 2-3 pulses duration using an electroporation buffer for cell
cultures). In addition, we confirmed that the above electroporation parameters
could not affect the choriogenic developmental program of follicles during
incubation in Grace’s medium. Next, the impact of systematic cis-element
mutations on the A/B.L9 chorion gene promoter were tested.
A revised model is proposed for the developmentally accurate regulation of the
A/B.L9 chorion genes. This model comprises a strict binding “program” of the
previously mentioned transcription factors on cis-regulatory elements, whose
access is dependent on two nucleosomes located on the promoter.
Keywords:
Gene expression, Bidirectional promoters, Choriogenesis, Electroporation, Bombyx mori
Number of references:
152
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