Shotgun sequencing of the human transcriptome with ORF expressed sequence tags

Shotgun sequencing of the human transcriptome with ORF expressed sequence tags

Author Dias Neto, Emmanuel Google Scholar
Correa, Ricardo Garcia Google Scholar
Verjovski-Almeida, Sergio Google Scholar
Briones, Marcelo Ribeiro da Silva Autor UNIFESP Google Scholar
Nagai, Maria Aparecida Google Scholar
Silva Junior, Wilson da Google Scholar
Zago, Marco Antonio Google Scholar
Bordin, Silvana Google Scholar
Costa, Fernando Ferreira Autor UNIFESP Google Scholar
Goldman, Gustavo Henrique Google Scholar
Carvalho, Alex Fiorini de Google Scholar
Matsukuma, Adriana Google Scholar
Baia, Gilson Soares Google Scholar
Simpson, David H. Google Scholar
Brunstein, Adriana Autor UNIFESP Google Scholar
Oliveira, Paulo Sergio Lopes de Google Scholar
Bucher, Philipp Google Scholar
Jongeneel, C. Victor Google Scholar
O'Hare, Michael J. Google Scholar
Soares, Fernando Google Scholar
Brentani, Ricardo Renzo Autor UNIFESP Google Scholar
Reis, Luis Fernando Lima Google Scholar
Souza, Sandro José de Google Scholar
Simpson, Andrew JG Google Scholar
Institution Ludwig Inst Canc Res
Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Fac Med Ribeirao Preto
Universidade Estadual de Campinas (UNICAMP)
Fac Ciencias Farmaceut Ribeirao Preto
Swiss Inst Bioinformat
Swiss Inst Expt Canc Res
Hosp AC Camargo Fund Antonio Prudente
Abstract Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome.
Language English
Date 2000-03-28
Published in Proceedings Of The National Academy Of Sciences Of The United States Of America. Washington: Natl Acad Sciences, v. 97, n. 7, p. 3491-3496, 2000.
ISSN 0027-8424 (Sherpa/Romeo, impact factor)
Publisher Natl Acad Sciences
Extent 3491-3496
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000086195200096

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