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Genetica ; 138(5): 519-30, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-19847655

RESUMO

The mariner-like transposon Mos1 is used for insertional mutagenesis and transgenesis in different animals (insects, nematodes), but has never been used in plants. In this paper, the transposition activity of Mos1 was tested in Nicotiana tabacum, but no transposition event was detected. In an attempt to understand the absence of in planta transposition, Mos1 transposase (MOS1) was produced and purified from transgenic tobacco (HMNtMOS1). HMNtMOS1 was able to perform all transposition reaction steps in vitro: binding to ITR, excision and integration of the same pseudo-transposon used in in planta transposition assays. The in vitro transposition reaction was not inhibited by tobacco nuclear proteins, and did not depend on the temperature used for plant growth. Several hypotheses are proposed that could explain the inhibition of HMNtMOS1 activity in planta.


Assuntos
Proteínas de Ligação a DNA/genética , Nicotiana/enzimologia , Nicotiana/genética , Transposases/genética , Sequência de Bases , Núcleo Celular/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Técnicas In Vitro , Microscopia de Fluorescência/métodos , Modelos Genéticos , Dados de Sequência Molecular , Plantas/genética , Plantas Geneticamente Modificadas , Ligação Proteica , Protoplastos/metabolismo , Proteínas Recombinantes/genética , Temperatura
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