Functional Identification of the Plasmodium Centromere and Generation of a Plasmodium Artificial Chromosome

Cell Host Microbe. 2010 Mar 18;7(3):245-55. doi: 10.1016/j.chom.2010.02.010.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Centromere*
  • Chromosomes
  • Chromosomes, Artificial*
  • Cloning, Molecular
  • Genetic Engineering / methods
  • Genetic Vectors*
  • Genetics, Microbial / methods
  • Plasmodium berghei / genetics*