Plasmid vectors and molecular building blocks for the development of genetic manipulation tools for Trypanosoma cruzi

PLoS One. 2013 Oct 24;8(10):e80217. doi: 10.1371/journal.pone.0080217. eCollection 2013.

Abstract

The post genomic era revealed the need for developing better performing, easier to use and more sophisticated genetic manipulation tools for the study of Trypanosoma cruzi, the etiological agent of Chagas disease. In this work a series of plasmids that allow genetic manipulation of this protozoan parasite were developed. First of all we focused on useful tools to establish selection strategies for different strains and which can be employed as expression vectors. On the other hand molecular building blocks in the form of diverse selectable markers, modifiable fluorescent protein and epitope-tag coding sequences were produced. Both types of modules were harboured in backbone molecules conceived to offer multiple construction and sub-cloning strategies. These can be used to confer new properties to already available genetic manipulation tools or as starting points for whole novel designs. The performance of each plasmid and building block was determined independently. For illustration purposes, some simple direct practical applications were conducted.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology
  • DNA, Protozoan
  • Genetic Engineering / methods*
  • Genetic Vectors*
  • Plasmids / genetics
  • Transfection
  • Trypanosoma cruzi / genetics*

Substances

  • DNA, Protozoan

Grants and funding

This work was supported by Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, PIP 2010-0685 and 2011-0263), Agencia Nacional de Promoción Científica y Tecnológica (FONCYT PICT 2008-1209 and 2010-0289), and Fundación Bunge y Born (grant 2012). CAP and MRM are members of the career of scientific investigator and MMC and LAB are research fellows from CONICET. LAB's research visit was financed by the E.P. Abraham Cephalosporin Fund and a Wood-Whelan Research Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.