Revista Cubana de Medicina Tropical
versão On-line ISSN 1561-3054
MONTALVO ALVAREZ, Ana Margarita; FOLGUEIRA FERNANDEZ, Cristina e REQUENA ROLANIA, José María. Cloning of 20 kDa heat shock protein of Leishmania amazonensis. Rev Cubana Med Trop [online]. 2009, vol.61, n.2, pp. 0-0. ISSN 1561-3054.
INTRODUCTION: the induction of heat shock proteins is a homeostatic mechanism that protects cells from the deleterious effects of thermal and other environmental stresses. In addition, they have important cell functions. The 20kDa heat shock protein in Leishmania amazonensis was recently reported. OBJECTIVE: to describe the methodology used for cloning of heat shock proteins, which allowed the study of some biological properties. METHODS: the hsp20 gene coding region was amplified by polymerase chain reaction using adequate primers. The amplified product was initially cloned in pCR2.1 vector (Invitrogen) and then in pET-28b vector (Novagen), to obtain recombinant protein. The same fragment was cloned also in the eukariote expression vector pcDNA3 (Invitrogen). The nucleotidic sequencing of the different clones was made, in order to verify their fidelity. RESULTS: the recombinant plasmids that encode HSP20 protein in Leishmania and allow obtaining massively recombinant protein and DNA were produced. CONCLUSIONS: both plasmids were useful to study some of the biological properties of this protein. This approach could be useful for similar research and represent a suitable methodological guideline.
Palavras-chave : heat shock proteins; Leishmania amazonensis; cloning; expression vector.