Molecular characterization of nitrate reductase gene and it expression in the marine red alga Gracilaria tenuistipitata (Rhodophyta) / Aspectos moleculares de nitrato redutase da macroalga marinha Gracilaria tenuistipitata (RHODOPHYTA): seqüenciamento do gene e estudo da expressão do RNA mensageiro

AUTOR(ES)

Vanessa dos Reis Falcão

DATA DE PUBLICAÇÃO

2006

RESUMO

In the marine environment, the major source of nitrogen is nitrate, which must be reduced before incorporation into organic compounds, such as amino acids and nitrogen bases (purines and pyrimidines). The first enzyme that participates in the nitrogen assimilation process is nitrate reductase (NR), which is responsible for the conversion of nitrate to nitrite. Gracilaria tenuistipitata is a marine macroalga with a great economic importance, harvested for the production of agar, which has many uses in products from industry and biotechnology. Nitrogen is the limiting macronutrient of this algas growth. The previous biochemical characterization for NR in this alga demonstrated a dial oscillation of activity and protein expression. These rhythms have been conserved in the situation of continuous light being regulated by the biological clock. For a better understanding of the regulation of NR, we sequenced the NR gene and studied the genetic regulation of NR mRNA expression. In a cDNA library of G. tenuistipitata, we identified and sequenced the NR clones, using the primer walking methodology. Using the same primers we also sequenced the gene that did not show introns. This is the first description in the literature of a NR gene for red algae and also of intron absence in NR genes. A rhythm for NR mRNA expression was observed in a period of 24h. In conclusion NR expression is also regulated at the transcription level. There is a basal expression of the NR gene that could be related to two hypotheses: two NR genes, one constitutive and another regulated, or only one NR gene, which is continuously translated having, however, its expression regulated and enhanced.

ASSUNTO(S)

nitrate reductase ritmo biológico seqüênciamento e expressão gênica red macroalgae macroalga vermelha biological rhythm gracilaria tenuistipitata gene sequence and expression gracilaria tenuistipitata nitrato redutase

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