TY - JOUR
T1 - A new method for isolation of purified genomic DNA from haemosporidian parasites inhabiting nucleated red blood cells
AU - Palinauskas, Vaidas
AU - Krizanauskiene, Asta
AU - Iezhova, Tatjana A.
AU - Bolshakov, Casimir V.
AU - Jönsson, Jane
AU - Bensch, Staffan
AU - Valkiunas, Gediminas
PY - 2013
Y1 - 2013
N2 - During the last 10 years, whole genomes have been sequenced from an increasing number of organisms. However, there is still no data on complete genomes of avian and lizard Plasmodium spp. or other haemosporidian parasites. In contrast to mammals, bird and reptile red blood cells have nuclei and thus blood of these vertebrates contains high amount of host DNA; that complicates preparation of purified template DNA from haemosporidian parasites, which has been the main obstacle for genomic studies of these parasites. In the present study we describe a method that generates large amount of purified avian haemosporidian DNA. The method is based on a unique biological feature of haemosporidian parasites, namely that mature gametocytes in blood can be induced to exflagellate in vitro. This results in the development of numerous microgametes, which can be separated from host blood cells by simple centrifugation. Our results reveal that this straight forward method provides opportunities to collect pure parasite DNA material, which can be used as a template for various genetic analyses including whole genome sequencing of haemosporidians infecting birds and lizards. (c) 2012 Elsevier Inc. All rights reserved.
AB - During the last 10 years, whole genomes have been sequenced from an increasing number of organisms. However, there is still no data on complete genomes of avian and lizard Plasmodium spp. or other haemosporidian parasites. In contrast to mammals, bird and reptile red blood cells have nuclei and thus blood of these vertebrates contains high amount of host DNA; that complicates preparation of purified template DNA from haemosporidian parasites, which has been the main obstacle for genomic studies of these parasites. In the present study we describe a method that generates large amount of purified avian haemosporidian DNA. The method is based on a unique biological feature of haemosporidian parasites, namely that mature gametocytes in blood can be induced to exflagellate in vitro. This results in the development of numerous microgametes, which can be separated from host blood cells by simple centrifugation. Our results reveal that this straight forward method provides opportunities to collect pure parasite DNA material, which can be used as a template for various genetic analyses including whole genome sequencing of haemosporidians infecting birds and lizards. (c) 2012 Elsevier Inc. All rights reserved.
KW - Avian malaria
KW - Haemosporidians
KW - Haemoproteus tartakovskyi
KW - hSISKIN1
KW - Template
KW - DNA purification
KW - Next generation sequencing
U2 - 10.1016/j.exppara.2012.12.003
DO - 10.1016/j.exppara.2012.12.003
M3 - Article
C2 - 23268173
SN - 0014-4894
VL - 133
SP - 275
EP - 280
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 3
ER -