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Identifier

000364436

Title

Establishing a genetic tool for mosaic analysis in the red flour beetle T. castaneum

Alternative Title

Η καθιέρωση ενός γενετικού εργαλείου για μωσαϊκή ανάλυση στο σκαθάρι T. castaneum

Author

Καλογεροπούλου Ελπινίκη

Thesis advisor

Αβέρωφ, Μιχάλης

Reviewer

Δεληδάκης, Χρήστος
Τζαμαρίας, Δημήτρης

Abstract

In recent years, new genetic tools have been established that enhance our ability to study in detail gene function, neuronal networks and cell and organism architecture across a range of different species. In this MSc project I focus on establishing the genetic tool brainbow in the emerging arthropod model Tribolium castaneum. Brainbow is a construct carrying 4 different fluorescent proteins and variants of lox sites flanking each one of them. In a transgenic animal carrying both a brainbow and a cre recombinase expressing construct, the lox sites recombine stochastically. This results in clones of cells, genetically labelled with different fluorescent proteins. First introduced in mice by Livet et al. 2007, brainbow was targeted to the CNS. I have made the first attempt to target brainbow expression ubiquitously in Tribolium castaneum. If successful, cells should be marked with different fluorescing proteins, such that their progeny can be tracked in vivo, enabling us to investigate how the growth zone elongates and segments form in Tribolium castaneum. I generated two different types of transgenic lines, some carrying a construct expressing cre recombinase (eight lines) and others carrying the brainbow construct (three lines). cre and brainbow lines were crossed with each other and then tested by PCR and imaging for efficient recombination of the constructs. PCR experiments suggest that the cre/lox system is working in vivo in Tribolium castaneum. Recombination between the lox variants is happening. In addition, confocal microscopy revealed cherry protein labelled clones, resulting from recombination between one of the pairs of lox sites. However, the observed clones are few, rare and very small. Results show that the heat shock promoter used to mediate cre recombinase expression is leaky.

Language

English

Subject

Brainbow

Cre/lox

Multicolour imaging

Transgenesis

Tribolium castaneum

Issue date

2011-03-18

Collection

School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses