Relevance and limitations of public databases for microarray design: a critical approach to gene predictions.
In conjunction with the completion of the human genome sequence, microarray technology offers a complementary strategy to traditional methodologies used to search for genetic determinants involved in multifactorial diseases such as Alzheimer's disease. In order to gain benefits from this strategy, we have designed home-made microarrays to compare the expression of all ORFs located within loci of interest defined by genome scanning in Alzheimer family studies. Two approaches were selected using either probes amplified by PCR from a cDNA bank or specific oligonucleotides. Here, we report the challenging task of validating, prioritising and selecting the best ORFs derived from the genome sequence. The initial inventory from the NCBI website allowed us to select 5849 ORF's within nine loci. Half of them resulted from prediction models using the GenomeScan software. However, our data have shown that predicted ORFs may not be representative of exonic sequences, or even real genes. These observations have led us to exclude these ORFs from our study, decreasing their number from 5849 to 2748. Microarrays may be only 'snapshots' of our current knowledge of the human genome.
Lambert
J C
JC
1Unité INSERM 508, Institut Pasteur de Lille, 1 rue du professeur Calmette, Lille cédex, France. jean-charles.lambert@Pasteur-lille.fr
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D018076:DNA, Complementary; D019992:Databases as Topic; D015894:Genome, Human; D006801:Humans; D008957:Models, Genetic; D009316:National Institutes of Health (U.S.); D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D014481:United States
D018076:DNA, Complementary