Functional identification of the Plasmodium centromere and generation of a Plasmodium artificial chromosome

Iwanaga, S., Khan, S.M., Kaneko, I., Christodoulou, Z., Newbold, C.N., Yuda, M., Janse, C.J. and Waters, A.P. (2010) Functional identification of the Plasmodium centromere and generation of a Plasmodium artificial chromosome. Cell Host and Microbe, 7(3), pp. 245-255. (doi:10.1016/j.chom.2010.02.010)

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Abstract

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.

Item Type:Articles (Other)
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Waters, Professor Andy
Authors: Iwanaga, S., Khan, S.M., Kaneko, I., Christodoulou, Z., Newbold, C.N., Yuda, M., Janse, C.J., and Waters, A.P.
Subjects:Q Science > QR Microbiology
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Cell Host and Microbe
ISSN:1931-3128
ISSN (Online):1934-6069

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
466861Conditional translational repression: a core regulatory mechanism of gene expression during development of the malaria parasite.Andrew WatersWellcome Trust (WELLCOME)083811/Z/07/ZInfection Immunity and Inflammation Life Sciences