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Cloning of a cDNA encoding DNA topoisomerase I in Daucus carota and expression analysis in relation to cell proliferation

Gene, ISSN: 0378-1119, Vol: 183, Issue: 1, Page: 183-190
1996
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Article Description

DNA topoisomerase I is an enzyme involved in several processes related to DNA metabolism. Despite the physiological importance, the regulation of topl gene expression has not yet been investigated in plants. In order to monitor the possible correlation between levels of topl transcripts and the proliferative state of the cell, two partially overlapping cDNAs encoding DNA topoisomerase I from Daucus carota have been isolated from a poly(A) + -primed library, using an Arabidopsis thaliana probe, and from a cDNA library spanning the 5′ region of the topl transcript, which was constructed using an antisense specific oligonucleotide. The topl nucleotide sequence encoded an open reading frame of 2370 bp, predicting a protein of 90 kDa. The deduced amino acid sequence showed a similarity of 51% with A. thaliana, 41% with S. cerevisiae, 40% with S. pombe and 31% with H. sapiens, respectively. Southern blot analysis, performed under moderate stringency conditions, showed the presence of a single-copy gene. Evaluation of the topl mRNA steady-state level revealed, besides a constitutive expression in vegetative carrot tissues, an induced expression related to cell proliferation.

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