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Closure of rRNA related gaps in the Chromobacterium violaceum genome with the PCR-assisted contig extension (PACE) protocol
Carraro, D. M; Camargo, A. A; Salim, A. C; Gonzaga, L; Costa, G. C; Vasconcelos, A. T; Simpson, A. J.
Afiliação
  • Carraro, D. M; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Camargo, A. A; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Salim, A. C; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Gonzaga, L; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Costa, G. C; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Vasconcelos, A. T; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
  • Simpson, A. J; Ludwig Institute for Cancer Research. Laboratory of Genetics São Paulo. BR
Genet. mol. res. (Online) ; 3(1): 53-63, Mar. 2004.
Article em En | LILACS | ID: lil-417585
Biblioteca responsável: BR1.1
ABSTRACT
In the finishing phase of the Chromobacterium violaceum genome project, the shotgun sequences were assembled into 57 contigs that were then organized into 19 scaffolds, using the information from shotgun and cosmid clones. Among the 38 ends resulting from the 19 scaffolds, 10 ended with sequences corresponding to rRNA genes (seven ended with the 5S rRNA gene and three ended with the 16S rRNA gene). The 28 non-ribosomal ends were extended using the PCR-assisted contig extension (PACE) methodology, which immediately closed 15 real gaps. We then applied PACE to the 16S rRNA gene containing ends, resulting in eight different sequences that were correctly assembled within the C. violaceum genome by combinatory PCR strategy, with primers derived from the non-repetitive genomic region flanking the 16S and 5S rRNA gene. An oriented combinatory PCR was used to correctly position the two versions (copy A and copy B, which differ by the presence or absence of a 100-bp insert); it revealed six copies corresponding to copy A, and two to copy B. We estimate that the use of PACE, followed by combinatory PCR, accelerated the finishing phase of the C. violaceum genome project by at least 40 per cent
Assuntos
Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: RNA Ribossômico / Reação em Cadeia da Polimerase / Chromobacterium / Genoma Bacteriano Idioma: En Revista: Genet. mol. res. (Online) Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2004 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil
Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: RNA Ribossômico / Reação em Cadeia da Polimerase / Chromobacterium / Genoma Bacteriano Idioma: En Revista: Genet. mol. res. (Online) Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2004 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil