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 | Acesso ao texto completo restrito à biblioteca da Embrapa Hortaliças. Para informações adicionais entre em contato com cnph.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Hortaliças. |
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Data corrente: |
05/12/2007 |
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Data da última atualização: |
24/01/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
FERREIRA, P. de T de O.; LEMOS, T. O.; NAGATA, T.; INOUE-NAGATA, A. K. |
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Afiliação: |
PAULO DE TARSO DE OLIVEIRA FERREIRA; THAÍS OLIVEIRA LEMOS, UNIVERSIDADE CATÓLICA DE BRASÍLIA; TATSUYA NAGATA, UNIVERSIDADE CATÓLICA DE BRASÍLIA; ALICE KAZUKO INOUE NAGATA, CNPH. |
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Título: |
One-step cloning approach for construction of agroinfectious begomovirus clones. |
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Ano de publicação: |
2008 |
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Fonte/Imprenta: |
Journal of Virological Methods, Amsterdam, v. 147, n. 2, p. 351-354, Feb. 2008. |
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ISSN: |
1879-0984 |
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DOI: |
10.1016/j.jviromet.2007.10.001 |
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Idioma: |
Inglês |
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Conteúdo: |
Most begomoviruses have a bipartite genome containing two circular ssDNA segments (DNA-A and DNA-B). Routine infectious clone construction relies upon cloning of the whole genome, which is then subcloned as a tandem one-and-half or two genome- (containing two replication origins) cassette into a vector prior to agro-inoculation. The construction of cassettes containing two replication origins is, however, a time-consuming process. Here an improved method for rapid construction of agroinfectious begomovirus clones is described. Total DNA was extracted from a tomato plant infected with Tomato golden vein virus and viral ssDNA molecules were amplified using phi-29 bacteriophage polymerase. High molecular weight DNA was partially digested with a single cutting restriction endonuclease (BamHI) and DNA fragments containing genome dimers were ligated into pCAMBIA0380, and used to transform Escherichia coli cells. This transformation yielded clones containing either DNA-A or DNA-B dimers. One clone each was used to transform Agrobacterium tumefaciens cells. DNA-A and DNA-B transformants were mixed and inoculated into test plants. All inoculated plants (tomato and Nicotiana benthamiana) became infected, confirming the infectivity of the clones. This approach was proven to be extremely fast and useful for the production of infectious clones. |
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Palavras-Chave: |
Cultivar Santa Clara. |
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Thesagro: |
Agrobacterium Tumefaciens; Clonagem; Tomate. |
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Thesaurus Nal: |
Begomovirus; Cloning (plants); Solanum lycopersicum. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02164naa a2200265 a 4500
001 1780604
005 2024-01-24
008 2008 bl uuuu u00u1 u #d
022 $a1879-0984
024 7 $a10.1016/j.jviromet.2007.10.001$2DOI
100 1 $aFERREIRA, P. de T de O.
245 $aOne-step cloning approach for construction of agroinfectious begomovirus clones.$h[electronic resource]
260 $c2008
520 $aMost begomoviruses have a bipartite genome containing two circular ssDNA segments (DNA-A and DNA-B). Routine infectious clone construction relies upon cloning of the whole genome, which is then subcloned as a tandem one-and-half or two genome- (containing two replication origins) cassette into a vector prior to agro-inoculation. The construction of cassettes containing two replication origins is, however, a time-consuming process. Here an improved method for rapid construction of agroinfectious begomovirus clones is described. Total DNA was extracted from a tomato plant infected with Tomato golden vein virus and viral ssDNA molecules were amplified using phi-29 bacteriophage polymerase. High molecular weight DNA was partially digested with a single cutting restriction endonuclease (BamHI) and DNA fragments containing genome dimers were ligated into pCAMBIA0380, and used to transform Escherichia coli cells. This transformation yielded clones containing either DNA-A or DNA-B dimers. One clone each was used to transform Agrobacterium tumefaciens cells. DNA-A and DNA-B transformants were mixed and inoculated into test plants. All inoculated plants (tomato and Nicotiana benthamiana) became infected, confirming the infectivity of the clones. This approach was proven to be extremely fast and useful for the production of infectious clones.
650 $aBegomovirus
650 $aCloning (plants)
650 $aSolanum lycopersicum
650 $aAgrobacterium Tumefaciens
650 $aClonagem
650 $aTomate
653 $aCultivar Santa Clara
700 1 $aLEMOS, T. O.
700 1 $aNAGATA, T.
700 1 $aINOUE-NAGATA, A. K.
773 $tJournal of Virological Methods, Amsterdam$gv. 147, n. 2, p. 351-354, Feb. 2008.
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Embrapa Hortaliças (CNPH) |
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| Registros recuperados : 4 | |
| 3. |  | ALBUQUERQUE, L. C.; VARSANI, A.; FERNANDES, F. R.; PINHEIRO, B; MARTIN, D. P.; FERREIRA, P. de T. O.; LEMOS, T. O.; INOUE NAGATA, A. K. Further characterization of tomato-infecting begomovíruses in Brazil. Archives of Virology, New York, v. 157, n. 4, p. 747-752, Apr. 2012. 0304-8608| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Hortaliças. |
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| 4. | | KITAJIMA, E. W.; KUBO, K. S.; FERREIRA, P. de T. O.; ALCÂNTARA, B. K. de; BOARI, A. J.; GOMES, R. T.; FREITAS-ÁSTUA, J.; REZENDE, J. A. M.; MORAIS, G. J. de; SALAROLI, R. B. Chlorotic spots on Clerodendrum, a disease caused by a nuclear type of Brevipalpus (Acari: Tenuipalpidae) transmitted virus. Scientia Agricola, Piracicaba, v. 6, n. 1, p. 36-49, jan/fev. 2008.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: Nacional - A |
| Biblioteca(s): Embrapa Mandioca e Fruticultura. |
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| Registros recuperados : 4 | |
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