Estudo dos polimorfismos no gene da NTPDase 2 da cepa ET de Leishmania (Viannia) braziliensis

• Autor(a) principal: Valiate, Bruno Vinicius Santos
• Data de Publicação: 2012
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: LOCUS Repositório Institucional da UFV
• Texto Completo: http://locus.ufv.br/handle/123456789/2454
• topic: Polimorfismo; NTPDase 2; Leishmania; Polymorphisms; NTPDase 2; Leishmania; CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR

Leishmaniasis are negligenciate diseases of sub-tropical countries. They are caused by protozoa that belong to Leishmania genus and are trasmitted by sand flies. Leishmaniasis mainly exists at cutaneous and visceral forms. At the Leishmania s surface there are proteins named ecto-triphosphate diphosphohydrolase (apyrases). Apyrases are enzymes that hydrolyze tri and di-phosfate nucleosides. Leishmania braziliensis has two mapped apyrase genes in its genome, NTPDase-1 and NTPDase-2. There are several papers that correlate the enzymatic activity with some processes like virulence, infection, nutrients capitation, and cellular adhesion. Non published data showed that there are single nucleotides polymorphisms (SNPs) in the messenger sequence of the NTPDase. SNPs might influence gene function by changing a codon base and its encoding amino acid. The present work proposed an analysis of SNPs from NTPDase-2 of ET strain of L.b. At the first part of the work, the genomic DNA was extracted to be used as a mold to the isolation of the NTPDase-2 gene by PCR. At the second part, clones of cultivated parasites were isolated through the limited dilution. After the isolation of five clones, the same procedure of the first part was made with the aim to sequence the NTPDase-2 gene of each isolate clone.A growth curve and measure of ecto-nucleotidase activity with the clones was made. The sequencing showed that in all the isolated colonies (in both at the first as at the second part) there are just 2 SNPs that do not change the codon. Two of five clones had a smaller growth. In the activity, the clones with the biggest growth had the biggest activity, it confirmed other results relating parasite`s proliferation and the ecto-nucleotidase activity. One of the clones, D12, had a high activity but did not grow so much. With this results, we can`t confirm the total relation but there are many mechanisms involving the cell growth and it may be involved with the difference between the activity and the clone D12 growth.