Max-Planck-Institut für Immunbiologie
Dept. Developmental Biology
telephone: +49 761 5108 582
fax: +49 761 5108 569
presenter: Bernhard Herrmann
Lorenz Neidhardt, Stephan Gasca, Karin Wertz, Andrea Schepler, Franz Obermayr, Alexander Aulehla, Susanne Worpenberg and Bernhard G. Herrmann
The molecular analysis of cell differentiation, tissue patterning and organ
development requires the isolation of genes involved in these processes. Large
scale EST and genome sequencing have been employed by many laboratories in order
to identify all genes encoded by the human genome. However, sequence information
alone does not reveal the tissue type, process or organ in which a particular
gene is acting.
We have developed a high-throughput strategy for determining the expression
of genes in 9.5 day mouse embryos on a large scale. Genes are selected from
cDNA libraries or cDNA arrays and assayed for their expression patterns by multiple
whole mount in situ hybridisation analyses, thus providing expression data at
high resolution. A genome set of 40,000 genes could be assayed in 20 person
years (i.e. 5 persons, 4 years). Gene expression and sequence information together
represent essential information about the cell type and process a gene is acting
in, and often also about the biochemical function. This information can be utilized
to develop strategies for further functional analyses, and to identify potential
target genes for pharmaceutical drugs. In addition, chromosomal mapping of the
genes identifies candidates for known human disorders and mouse mutations.
In a pilot screen, appr. 800 specifically expressed genes have been identified.
Of 607 clones sequenced, about 43% represent known genes and about 44% novel
genes or ESTs. The remaining cDNAs represent mouse orthologues of known genes
or new members of known gene families.
Appr. 25% of the known genes isolated are involved in transcriptional regulation, such as Hox genes, Tbx genes, Msx, Pax9. Appr. 16% of the known genes isolated represent mouse orthologues of human disorders. Three mouse genes have been isolated for which a classical mutant locus is known (Brachyury, pudgy, vibrator). This suggests that a similar proportion of mouse and human mutations is represented among the novel genes and ESTs identified. A data base providing expression, sequence and homology search data is being built up at the Resource Center (RZPD). This project will have a strong impact on several fields of biomedical research.
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