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Colombia Médica
versão On-line ISSN 1657-9534
Resumo
TORO-LONDONO, Miguel A. et al. Leishmania (Viannia) panamensis expresa una nucleasa molecular y bioquímicamente similar a la Endonucleasa G de eucariotas superiores. Colomb. Med. [online]. 2011, vol.42, n.2, pp.154-165. ISSN 1657-9534.
Objective: To characterize the molecular and biochemical features of the Endonuclease G of Leishmania (Viannia) panamensis. Methods: The gene of the putative L. (V.) panamensis Endonuclease G was amplified, cloned, and sequenced. The recombinant protein was produced in a heterologous expression system and biochemical assays were run to determine its ion, temperature, and pH preferences. Results: The L. (V.) panamensis rENDOG has biochemical features similar to those found in other trypanosomatids and higher eukaryotes. In addition, phylogenetic analysis revealed a possible evolutionary relationship with metazoan ENDOG. Conclusions: L. (V.) panamensis has a gene that codifies an ENDOG homologous to those of higher organisms. This enzyme can be produced in Escherichia coli and is able to degrade covalently closed circular double-stranded DNA. It has a magnesium preference, can be inhibited by potassium, and is able to function within a wide temperature and pH range.
Palavras-chave : Leishmania panamensis; Endonucleasa G; Caracterización molecular; Proteínas recombinantes; Renaturalización; Filogenia.