Isolation and characterisation of a cDNA encoding a novel cytosolic ascorbate peroxidase from potato plants (Solanum tuberosum L.) |
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Authors: | Diana Soares da Costa Cláudia Sofia Pereira Jorge Teixeira Susana Pereira |
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Institution: | (1) Plant Functional Biology Department, Institute for Cell and Molecular Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal;(2) Botany Department, School of Sciences, University of Porto Rua do Campo Alegre, 1191, 4150-181 Porto, Portugal |
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Abstract: | Ascorbate peroxidase (APX) is one of the key enzymes of the plant antioxidant system playing, along with catalase, a central
role in hydrogen peroxide scavenging. An approach to further increase the knowledge about cytosolic APX gene organization
can be achieved by isolating and characterisating new cDNAs, thus providing new insights about the physiological roles and
regulation of these enzymes. A partial cDNA clone (corresponding to the 3’ untranslated region), cytosolic ascorbate peroxidase-related,
was isolated from potato sprouts by RT-PCR. Database analysis retrieved several expressed sequence tags (ESTs) coding potato
cytosolic ascorbate peroxidase, that were used to infer the complete cDNA sequence. The deduced amino acid sequence revealed
high homologies with other plant cytosolic ascorbate peroxidases, confirming the reliability of the virtual cDNA. Northern
blot analysis revealed the existence of a single band related to the isolated cDNA and the southern blotting results allowed
the elaboration of a possible gene organization. |
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Keywords: | ascorbate peroxidase cDNA isolation northern blotting potato southern blotting |
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