title: A new method for isolation of purified genomic DNA from haemosporidian parasites inhabiting nucleated red blood cells.
abstract: 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.
authors.affiliation: Nature Research Centre, Akademijos 2, Vilnius LT-08412, Lithuania. vaidas@ekoi.lt
authors.forename: Vaidas
authors.initials: V
authors.lastname: Palinauskas
authors.forename: Asta
authors.initials: A
authors.lastname: Križanauskienė
authors.forename: Tatjana A
authors.initials: TA
authors.lastname: Iezhova
authors.forename: Casimir V
authors.initials: CV
authors.lastname: Bolshakov
authors.forename: Jane
authors.initials: J
authors.lastname: Jönsson
authors.forename: Staffan
authors.initials: S
authors.lastname: Bensch
authors.forename: Gediminas
authors.initials: G
authors.lastname: Valkiūnas
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000097:blood / Q000469:parasitology*; D002467:Cell Nucleus / Q000235:genetics; D016054:DNA, Protozoan / Q000097:blood / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D004912:Erythrocytes / Q000469:parasitology*; D018503:Genome, Protozoan / Q000235:genetics; D016787:Haemosporida / Q000235:genetics*; D046109:Passeriformes / Q000097:blood / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011529:Protozoan Infections, Animal / Q000097:blood / Q000469:parasitology*; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D016054:DNA, Protozoan