Vertebrate exon trapping methods: implications for transcript mapping with mosquito DNA
Autor(es): Brown, S. E.; Wood, S. H.; Knudson, D. L.
Resumo: Exon trapping methods have played an important role in the development of transcript maps. In one in vivo vertebrate method, exons in a genomic DNA clone are transcribed, and they are recovered without any a priori information on the nature of the expressed transcript. The only requirement is that the genomic DNA clone contains exons separated by intervening introns that are removed by splicing during mRNA transcription and that the splice donor and acceptor site sequences follow those used by vertebrates. It is not known whether invertebrate splice donor and acceptor sites from genes that contain short introns will be processed correctly using an in vivo vertebrate exon trapping method. In this report, an analysis of mosquito splice sites using software designed to identify exons in genomic DNA sequence suggested that the vertebrate exon trapping method could recognize mosquito introns and exons. When a mosquito genomic DNA clone containing the D7 gene was tested experimentally, this method failed to recognize and process small introns (<63 bp) faithfully. In spite of this failure, exons and exon fragments were recovered. The implications of these findings and their application to map-based positional cloning in mosquito genomics is discussed. (C) 1999 Elsevier Science Ltd. All rights reserved.
Palavras-Chave: Bioinformatics; D7 gene; Exon trapping; Intron; RNA splicing; Transcript mapping
Imprenta: Insect Biochemistry and Molecular Biology, v. 29, n. 7, p. 643-651, 1999
Identificador do objeto digital: 10.1016/S0965-1748(99)00042-9
Descritores: Aedes aegypti - DNA ; Aedes aegypti - RNA
Data de publicação: 1999
