Uma investigaÃÃo: ESTs (expressed sequence tags) podem ser usados no desenvolvimento de marcadores moleculares baseados em introns?

AUTOR(ES)
DATA DE PUBLICAÇÃO

2003

RESUMO

Recent advances in Molecular Biology have allowed the development of powerful techniques for genetic diversity analysis. Use of the Polymerase Chain Reaction (PCR) has greatly facilitated the development of molecular markers for genetic mapping, and comparative mapping of different species. For comparative mapping it is essential to use coding regions of the genome. It is known that ESTs are the most rapid way to obtain information about coding sequences and, therefore, they could potentially be used as a tool for molecular marker development. The main purpose of this work was to investigate the possibility of molecular marker development based on introns from Arachis stenosperma and Arachis hypogaea ESTs (Expressed Sequence Tags). This work was done as part of a larger project âTHE IDENTIFICATION OF RESISTANCES TO BIOTIC STRESS IN WILD ARACHIS GERMPLASM, AND THE DEVELOPMENT OF TOOLS FOR BREEDING BY GENETIC MAPPING AND COMPARATIVE GENOMICS.â, which has as one of its purposes to identify, among wild peanut species, sources of resistance against South American pests and diseases. It has been shown that. A stenosperma is a wild South American species that has resistance against fungi and nematodes. Therefore, in the present work, through the production, sequencing and computer analysis of ESTs obtained from an A stenosperma leaf cDNA databank, we were able to develop a molecular marker candidate search methodology based on introns. ESTs from A.stenosperma root inoculated or not with Bradiryzobium, as well as ESTs from A. hypogaea obtained from dbEST databank, were also used in this study. In order to automate the marker searching process, a computational tool for searching for homologues with specific introns, and the alignment between them and their correspondent ESTs, was developed. From 330 ESTs of A.stenosperma, we were able to find 4 markers candidates based on introns, while from 1346 A. hypogaea, we identified 9 candidates. These markers were used to design primers flanking introns that can be tested in the general project. Therefore, through the preliminary results obtained from the present investigation, there is evidence that it is possible to develop molecular markers based on introns.

ASSUNTO(S)

ciÃncias biolÃgicas marcadores moleculares biologia computacional biologia molecular genetic diversity introns molecular biology ests arachis

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