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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.

Original publication

DOI

10.1016/j.chom.2010.02.010

Type

Journal article

Journal

Cell Host Microbe

Publication Date

18/03/2010

Volume

7

Pages

245 - 255

Keywords

Centromere, Chromosomes, Chromosomes, Artificial, Cloning, Molecular, Genetic Engineering, Genetic Vectors, Genetics, Microbial, Plasmodium berghei