Amplificación, clonación y expresión de los genes (kmp11, hsp70, lack ,tsa y hlyA) de leishmania en cepas recombinantes de escherichia coli

Abstract

The study, development and application of molecular techniques, especially recombinant DNA technology, has become an alternative for the production of antigens, acquiring great relevance due to the biological, chemical and medical utilities. The main objective of this project was the amplification, cloning and gene expression (kmp11, hsp70, tsa, lack and hlyA) of Leishmania infantum in recombinant strains of E.coli (MC4100, JC8100 and BL-21) bacteria. For this purpose, extraction protocols (plasmid extraction with Econospin column), transformation (thermal shock transformation [freezing - defrosting] - sonication, TSS solution and Electroporation) and amplification (Touchdown type PCR) were performed. The products of these procedures were analyzed by agarose gel electrophoresis, Dot Blot and Western Blot, which by means of bands manages to suppose the presence of the antigens of interest. It is expected that this procedure will be useful for the contribution to the analysis of immunological activity in living organisms and later manufacture of vaccines or treatments of leishmaniasis disease.