Journal
CELL HOST & MICROBE
Volume 7, Issue 3, Pages 245-255Publisher
CELL PRESS
DOI: 10.1016/j.chom.2010.02.010
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Funding
- Ministry of Education, Science, Culture, and Sports of Japan [21022031, 21790406, 20249023, 21022019]
- Wellcome Trust Post-Genome Initiative Grant
- Principal Research Fellowship
- European Network of Excellence, BioMalPar
- Grants-in-Aid for Scientific Research [21022031, 20249023, 21790406, 21022019] Funding Source: KAKEN
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The artificial chromosome represents a useful tool for gene transfer, both as cloning vectors and in chromosome biology research. To generate a Plasmodium artificial chromosome (PAC), we had to first functionally identify and characterize the parasite's centromere. A putative centromere (pbcen5) was cloned from chromosome 5 of the rodent parasite P. berghei based on a Plasmodium gene-synteny map. Plasmids containing pbcen5 were stably maintained in parasites during a blood-stage infection with high segregation efficiency, without drug pressure. pbcen5-containing plasmids were also stably maintained during parasite meiosis and mitosis in the mosquito. A linear PAC (L-PAC) was generated by integrating pbcen5 and telomere into a plasmid. The L-PAC segregated with a high efficiency and was stably maintained throughout the parasite's life cycle, as either one or two copies. These results suggest that L-PAC behaves like a Plasmodium chromosome, which can be exploited as an experimental research tool.
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