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Title: Shotgun sequencing of the human transcriptome with ORF expressed sequence tags
Authors: Dias Neto, Emmanuel
Correa, Ricardo Garcia
Verjovski-Almeida, Sergio
Briones, Marcelo Ribeiro da Silva [UNIFESP]
Nagai, Maria Aparecida
Silva Junior, Wilson da
Zago, Marco Antonio
Bordin, Silvana
Costa, Fernando Ferreira [UNIFESP]
Goldman, Gustavo Henrique
Carvalho, Alex Fiorini de
Matsukuma, Adriana
Baia, Gilson Soares
Simpson, David H.
Brunstein, Adriana [UNIFESP]
Oliveira, Paulo Sergio Lopes de
Bucher, Philipp
Jongeneel, C. Victor
O'Hare, Michael J.
Soares, Fernando
Brentani, Ricardo Renzo [UNIFESP]
Reis, Luis Fernando Lima
Souza, Sandro José de
Simpson, Andrew JG
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
Issue Date: 28-Mar-2000
Publisher: Natl Acad Sciences
Citation: 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.
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.
ISSN: 0027-8424
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