Expression and purification of the Leishmania major telomerase RT (reverse transcriptase)domain with the aim to perform interaction assays

• Grant number: 18/03095-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2018
• End date: December 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Maria Isabel Nogueira Cano
• Grantee: Vitor Luiz da Silva
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Leishmaniasis is a neglected tropical disease caused by parasites of the family Trypanosomatidae, genus Leishmania. The current treatment protocol implies highly toxic, high cost and can lead to resistant parasites. The absence of effective treatments to combat the disease, makes it important to seek for new therapeutic targets against Leishmania. Recently, studies about telomeres have been considered of great relevance due to its importance in the maintenance of genome stability and the parasite cellular proliferation. Telomeres are maintained by the action of a ribonucleoprotein, named telomerase, composed of the protein component, TERT (telomerase reverse transcriptase) and an RNA (TER), which contains the template telomeric sequence added at the end of the chromosomes. TERT protein bears structural domains that play different functions. In the present work we are initiating studies about the L. major RT (Reverse Transcriptase) domain which was cloned in a bacterial expression vector. Preliminary results show that the RT domain is expressed in the bacterial system and in the insoluble fraction of the bacterial extract. The aim of this project is to obtain an efficient protocol for its purification in order to perform in vitro protein-nucleic acid interaction assays.