Replication, expression and segregation of plasmid-borne DNA in genetically transformed malaria parasites.
To fully exploit the transfection technology developed for Plasmodium we investigated the features of replication, expression and segregation of an episomally maintained DNA construct during a sexual blood stage development in genetically transformed parasites of P. berghei. Using DNA in situ hybridisation techniques we were able to show that the introduced DNA construct is located in the nucleus of the parasite and is not segregating uniformly during schizogony. Replication of the construct mainly takes place between 16 and 24 h after invasion of the merozoites, coinciding with chromosomal replication. Furthermore the plasmid-borne DHFR/TS gene is constitutively transcribed throughout the asexual blood stage development. Hence the DHFR/TS promoter would appear to be a useful tool in the study of (over)expression of introduced genes and performing complementation studies in transfected parasites during the complete a sexual blood stage development of P. berghei.
van Dijk
M R
MR
Department of Parasitology, University of Leiden, The Netherlands. M.R.van_Dijk@Parasitology.MedFac.LeidenUniv.NL
Vinkenoog
R
R
Ramesar
J
J
Vervenne
R A
RA
Waters
A P
AP
Janse
C J
CJ
D000818:Animals; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D017403:In Situ Hybridization; D008288:Malaria / Q000235:genetics* / Q000469:parasitology*; D051379:Mice; D009097:Multienzyme Complexes / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D010962:Plasmodium berghei / Q000235:genetics*; D051381:Rats; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D013940:Thymidylate Synthase / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics*
D009097:Multienzyme Complexes; C056930:thymidylate synthase-dihydrofolate reductase; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase