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Prof. Dr. Ansgar Klebes
Freie Universität Berlin
Dept. of Genetics
Arnimallee 7
14195 Berlin, Germany

 

Fig. 1 Enlargement of 6 blocks of a DNA microarray hybridized with RNA probes from Drosophila adult flies (red channel) and embryos (green channel). Transcripts that are expressedat comparable levels in both samples appear yellow. Every block contains 540 spots of unique DNAs. Each spot represents 1 open reading frame of the Drosophila genome. With more than 17.000 (partially redundant) spots this array covers most of the predicted 14.000 (1st annotation) Drosophila genes.
 

research interest

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publications

Klebes A, Sustar A, Kechris K, Li H, Schubiger G, Kornberg TB. Regulation of cellular plasticity in Drosophila imaginal disc cells by the Polycomb group, trithorax group and lama genes. Development. 2005 Oct;132(19):4419.

Xu EY, Lee DF, Klebes A, Turek PJ, Kornberg TB, Reijo Pera RA. Human BOULE gene rescues meitoic defects in infertile flies. Hum Mol Gent. 2003 Jan 15;12(2):169-175.

Klebes A, Biehs B, Cifuentes F, Kornberg TB. Expression profiling of Drosophila imaginal discs. Genome Biol. 2002 Jul 24;3(8):0038

Abstract available below, full text free of charge. Click here to read full text: http://genomebiology.com/2002/3/8/research/0038

den Hollander AI, Johnson K, de Kok YJ, Klebes A, Brunner HG, Knust E, Cremers FP. CRB1 has a cytoplasmic domain that is functionally conserved between human and Drosophila. Hum Mol Genet. 2001 Nov 15;10(24):2767-73.

Bossinger O, Klebes A, Segbert C, Theres C, Knust E. Zonula adherens formation in Caenorhabditis elegans requires dlg-1, the homologue of the Drosophila gene discs large. Dev Biol. 2001 Feb 1;230(1):29-42.

Klebes A, Knust E. A conserved motif in Crumbs is required for E-cadherin localisation and zonula adherens formation in Drosophila. Curr Biol. 2000 Jan 27;10(2):76-85.

 

 

Expression profiling of Drosophila imaginal discs

Ansgar Klebes1, Brian Biehs1, Francisco Cifuentes1, 2 and Thomas B Kornberg1
1Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA
2Agilent Inc., 395 Page Mill Rd, Palo Alto, CA 94303, USA
Correspondence: Thomas B Kornberg. E-mail: tkornberg@biochem.ucsf.edu
Genome Biology 2002 3(8):research0038.1-0038.16 (published 24 July 2002)

Abstract

Background In the Drosophila larva, imaginal discs are programmed to produce adult structures at metamorphosis. Although their fate is precisely determined, these organs remain largely undifferentiated in the larva. To identify genes that establish and express the different states of determination in discs and larval tissues, we used DNA microarrays to analyze mRNAs isolated from single imaginal discs.

Results Linear amplification protocols were used to generate hybridization probes for microarray analysis from poly(A)+ RNA from single imaginal discs containing between 10,000 and 60,000 cells. Probe reproducibility and degree of representation were tested using microarrays with approximately 6,000 different cDNAs. Hybridizations with probes that had been prepared separately from the same starting RNA pool had a correlation coefficient of 0.97. Expression-profile comparisons of the left and right wing imaginal discs from the same larva correlated with a coefficient of 0.99, indicating a high degree of reproducibility of independent amplifications. Using this method, we identified genes with preferential expression in the different imaginal discs using pairwise comparisons of discs and larval organs. Whereas disc-to-disc comparisons revealed only moderate differences, profiles differed substantially between imaginal discs and larval tissues, such as larval endodermal midgut and mesodermal fat body.

Conclusion The combination of linear RNA amplification and DNA microarray hybridization allowed us to determine the expression profiles of individual imaginal discs and larval tissues and to identify genes expressed in tissue-specific patterns. These methods should be widely applicable to comparisons of expression profiles for tissues or parts of tissues that are available only in small amounts.

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last updated 2/13/06