TY - JOUR
T1 - Molecular characterization of the short interspersed repetitive element SIRE in the six discrete typing units (DTUs) of Trypanosoma cruzi
AU - Pavia, Paula X.
AU - Thomas, M. Carmen
AU - López, Manuel C.
AU - Puerta, Concepción J.
N1 - Funding Information:
The authors express gratitude to Dr. Michell Tybayrenc from Genetique et Evolution des Maladies Infectieuses (GEMI) and Dr. Octavio Fernández from the Oswaldo Cruz Institute for supplying the DNA from some T. cruzi isolates, to Marleny Montilla from the Parasitology Group, National Health Institute for supplying the cultures of some of the parasite isolates, and to Dr. Alejandro Schijman from Instituto de Investigaciones en Ingenieria Genetica y Biologia Molecular (INGEBI) for his assistance and advice in real time PCR analyses. This work was funded by the Vicerrectoria Académica at Pontificia Universidad Javeriana, project No. 2063. PXPV was funded by the National PhD Program of the Administrative Department of Science, Technology, and Innovation (Colciencias) and the Permanent Formation Program, 2008 from Fundación Carolina (Spain). MC López and MC Thomas were supported by Grants P08-CVI-04037 from PAI (Junta de Andalucía), BFU2010-1670 from Plan Nacional I+D+i (MICINN) and RD06/0021/0014 from ISCIII-RETIC (MICINN) Spain and FEDER.
PY - 2012/10
Y1 - 2012/10
N2 - Repetitive sequences constitute an important proportion of the Trypanosoma cruzi genome; hence, they have been used as molecular markers and as amplification targets to identify the parasite presence via PCR. In this study, a molecular characterization of the SIRE repetitive element was performed in the six discrete typing units (DTUs) of T. cruzi. The results evidenced that this element, located in multiple chromosomes, was interspersed in the genome of all DTUs of the parasite. The presence of several motifs implicated in element insertion, duplication, and functionality suggests that SIRE could be an active element in the parasite genome. Of interest, there were SIRE specific Alu I fragments that allowed to discriminate DTU I from the others DTUs. Moreover, an UPGMA phenetic tree constructed from fragment sharing Southern blot data showed that T. cruzi I isolates conform a cluster separated from the T. cruzi II-VI isolates. When the relative number of SIRE copies was determined, a variation from 105 to 2,000 copies per haploid genome was observed among the different isolates without kept a DTU-relationship. In all, these findings suggest that SIRE sequence is a good target for parasite DNA amplification.
AB - Repetitive sequences constitute an important proportion of the Trypanosoma cruzi genome; hence, they have been used as molecular markers and as amplification targets to identify the parasite presence via PCR. In this study, a molecular characterization of the SIRE repetitive element was performed in the six discrete typing units (DTUs) of T. cruzi. The results evidenced that this element, located in multiple chromosomes, was interspersed in the genome of all DTUs of the parasite. The presence of several motifs implicated in element insertion, duplication, and functionality suggests that SIRE could be an active element in the parasite genome. Of interest, there were SIRE specific Alu I fragments that allowed to discriminate DTU I from the others DTUs. Moreover, an UPGMA phenetic tree constructed from fragment sharing Southern blot data showed that T. cruzi I isolates conform a cluster separated from the T. cruzi II-VI isolates. When the relative number of SIRE copies was determined, a variation from 105 to 2,000 copies per haploid genome was observed among the different isolates without kept a DTU-relationship. In all, these findings suggest that SIRE sequence is a good target for parasite DNA amplification.
KW - Discrete typing units (DTUs)
KW - Short interspersed repetitive element (SIRE)
KW - TcH2AF-R PCR
KW - Trypanosoma cruzi
UR - http://www.scopus.com/inward/record.url?scp=84866407303&partnerID=8YFLogxK
U2 - 10.1016/j.exppara.2012.06.007
DO - 10.1016/j.exppara.2012.06.007
M3 - Artículo
C2 - 22750455
AN - SCOPUS:84866407303
SN - 0014-4894
VL - 132
SP - 144
EP - 150
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 2
ER -