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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Generation of longer 3'cDNA fragments from massively parallel signature sequencing tags

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Autor(es):
Silva, A. P. M. [1] ; Chen, J. ; Carraro, D. M. ; Wang, S. M. ; Camargo, A. A.
Número total de Autores: 5
Afiliação do(s) autor(es):
[1] Instituto Ludwig de Pesquisa sobre o Câncer. Laboratório de Biologia Molecular e Genômica - Brasil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: Nucleic Acids Research; v. 32, n. 12, p. e94-e97, jul. 2004.
Área do conhecimento: Ciências Biológicas - Genética
Assunto(s):Genética médica   Expressão gênica   Genomas
Resumo

Massively Parallel Signature Sequencing (MPSS) is a powerful technique for genome-wide gene expression analysis, which, similar to SAGE, relies on the production of short tags proximal to the 3'end of transcripts. A single MPSS experiment can generate over 107 tags, providing a 10-fold coverage of the transcripts expressed in a human cell. A significant fraction of MPSS tags cannot be assigned to known transcripts (orphan tags) and are likely to be derived from transcripts expressed at very low levels (1 copy per cell). In order to explore the potential of MPSS for the characterization of the human transcriptome, we have adapted the GLGI protocol (Generation of Longer cDNA fragments from SAGE tags for Gene Identification) to convert MPSS tags into their corresponding 3' cDNA fragments. GLGI-MPSS was applied to 83 orphan tags and 41 cDNA fragments were obtained. The analysis of these 41 fragments allowed the identification of novel transcripts, alternative tags generated from polymorphic and alternatively spliced transcripts, as well as the detection of artefactual MPSS tags. A systematic large-scale analysis of the genome by MPSS, in combination with the use of GLGI-MPSS protocol, will certainly provide a complementary approach to generate the complete catalog of human transcripts. (AU)

Processo FAPESP: 98/14335-2 - Antonio Prudente Cancer Research Center
Beneficiário:Fernando Augusto Soares
Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs