Genetics. Published Articles Ahead of Print: June 21, 2005, Copyright © 2005
doi:10.1534/genetics.105.041848


A more recent version of this article appeared on October 1, 2005.

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Minos as a genetic and genomic tool in Drosophila melanogaster

Athanasios Metaxakis 1, Stefan Oehler 2, Apostolos Klinakis 1 and Charalambos Savakis 3*

1 University of Crete and IMBB-FoRTH
2 IMBB-FoRTH
3 IMBB-FoRTH and University of Crete

* To whom correspondence should be addressed. E-mail: savakis{at}imbb.forth.gr.

Submitted on February 10, 2005
Revised on March 25, 2005
Accepted on 14 June 2005


Abstract

Much of the information about the function of D. melanogaster genes has come from P element mutagenesis. The major drawback of the P element, though, is its strong bias for insertion into some genes (hotspots) and against insertion into others (coldspots). Additionally, 5'UTRs are preferential intragenic targets. For the successful completion of the Drosophila Genome Disruption Project the use of transposon vectors other than P will be necessary. We have here examined the suitability of the Minos element from Drosophila hydei, which is a member of the Tc1/mariner family of transposable elements, as a tool for Drosophila genomics. Previous work has shown that Minos is active in diverse organisms and cultured cells; it produces stable integrants in the germ line of several insect species, in the mouse and in human cells. We generated and analyzed 96 Minos integrations into the Drosophila genome and devised an efficient "jumpstarting" scheme for production of single insertions. The ratio of insertions into genes versus intergenic DNA is consistent with a random distribution. Within genes, there is a statistically significant preference for insertion into introns rather than exons. About 30% of all insertions were in introns and 40% of intragenic insertions were in genes that have so far not been hit by the P-element. The insertion sites exhibit, in contrast to other transposons, very little sequence requirement beyond the TA dinucleotide insertion target. In addition, induced remobilization of Minos insertions can delete nearby sequences. Our results suggest that Minos is a useful tool complementing the P-element for insertional mutagenesis and genomic analysis in Drosophila.

Key Words: Drosophila gene disruption, Functional genomics, Insertional mutagenesis, Minos transposable element




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