Microbiologia

URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11840

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2002 - 20082

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Autor: search.filters.author.Lima, Juliana OliveiraData inicial: 2000Data final: 2009Assunto: search.filters.subject.Transformation

Resultados da Pesquisa

Agora exibindo 1 - 2 de 2
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    Restriction enzyme improves the efficiency of genetic transformations in Moniliophthora perniciosa, the causal agent of witches’ broom disease in Theobroma cacao
    (Brazilian Archives of Biology and Technology, 2008-01) Lopes, Francis Julio Fagundes; Queiroz, Marisa Vieira de; Lima, Juliana Oliveira; Silva, Viviane Aline Oliveira; Araújo, Elza Fernandes de
    The presence of restriction enzymes in the transformation mixture improved the efficiency of transformation in Moniliophthora perniciosa. The influence of the vector shape (linear or circular), the patterns of plasmid integration in genomic sites and the influence of the promoter used to express the gene marker were also analyzed. The addition of BamHI or NotI increased the number of transformants by 3-10-fold and 3-fold, respectively, over the control without added enzyme. The use of pre-linearized plasmid did not increase the transformation efficiency in comparison with the circular plasmid. However, the frequency of multi-copy transformants increased significantly. The transformation procedure here reported resulted in better production of protoplasts and transformation efficiency. In addition, the time necessary for the detection of the first transformants and the number of insertions were reduced.
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    Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants
    (Current Genetics, 2002-12-17) Lima, Juliana Oliveira; Santos, Jildete Karla dos; Pereira, Jorge Fernando; Resende, Mário Lúcio Vilela de; Araújo, Elza Fernandes de; Queiroz, Marisa Vieira de
    Protoplasts of the pathogenic plant fungus, Crinipellis perniciosa, were transformed to hygromycin B resistance using the pAN7-1 plasmid, which contains the Escherichia coli hph gene under the control of Aspergillus nidulans regulatory sequences. The pAN7-1 plasmid was introduced by PEG/CaCl2 treatment. Transformation frequencies of 1.6–2.5 transformants/μg of DNA were achieved. About 54% of the transformants were abortive and 40 analyzed transformants were mitotically stable and showed different hygromycin B resistance levels. The presence of the hph gene was checked by PCR in five transformants and the integration of multiple plasmid copies into different genome sites was observed by Southern analysis. This is the first report of a C. perniciosa transformation system and represents an important step for further research into genetic manipulation of this fungal plant pathogen.