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A new method for isolation of purified genomic DNA from haemosporidian parasites inhabiting nucleated red blood cells

Palinauskas, Vaidas; Krizanauskiene, Asta; Iezhova, Tatjana A.; Bolshakov, Casimir V.; Jönsson, Jane LU ; Bensch, Staffan LU and Valkiunas, Gediminas (2013) In Experimental Parasitology 133(3). p.275-280
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Avian malaria, Haemosporidians, Haemoproteus tartakovskyi, hSISKIN1, Template, DNA purification, Next generation sequencing
in
Experimental Parasitology
volume
133
issue
3
pages
275 - 280
publisher
Elsevier
external identifiers
  • wos:000316093100008
  • scopus:84872423541
ISSN
0014-4894
DOI
10.1016/j.exppara.2012.12.003
project
Malaria in birds
BECC
language
English
LU publication?
yes
id
d69e50c1-3034-49a6-8b42-9ceb19f9e656 (old id 3636574)
date added to LUP
2013-04-24 14:03:29
date last changed
2019-04-02 01:26:19
@article{d69e50c1-3034-49a6-8b42-9ceb19f9e656,
  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. (c) 2012 Elsevier Inc. All rights reserved.},
  author       = {Palinauskas, Vaidas and Krizanauskiene, Asta and Iezhova, Tatjana A. and Bolshakov, Casimir V. and Jönsson, Jane and Bensch, Staffan and Valkiunas, Gediminas},
  issn         = {0014-4894},
  keyword      = {Avian malaria,Haemosporidians,Haemoproteus tartakovskyi,hSISKIN1,Template,DNA purification,Next generation sequencing},
  language     = {eng},
  number       = {3},
  pages        = {275--280},
  publisher    = {Elsevier},
  series       = {Experimental Parasitology},
  title        = {A new method for isolation of purified genomic DNA from haemosporidian parasites inhabiting nucleated red blood cells},
  url          = {http://dx.doi.org/10.1016/j.exppara.2012.12.003},
  volume       = {133},
  year         = {2013},
}