A method for rapid genetic integration into Plasmodium falciparum utilizing mycobacteriophage Bxb1 integrase

Sophie H. Adjalley, Marcus C.S. Lee, David A. Fidock

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

41 Citations (Scopus)

Abstract

Genetic manipulation of the human malaria parasite Plasmodium falciparum has presented substantial challenges for research efforts aimed at better understanding the complex biology of this highly virulent organism. The development of methods to perform gene disruption, allelic replacement or transgene expression has provided important insights into the function of parasite genes. However, genomic integration studies have been hindered by low transfection and recombination efficiencies, and are complicated by the propensity of this parasite to maintain episomal replicating plasmids. We have developed a fast and efficient site-specific system of integrative recombination into the P. falciparum genome, which is catalyzed by the mycobacteriophage Bxb1 serine integrase. This system has the dvantage of providing greater genetic and phenotypic homogeneity within transgenic lines as compared to earlier methods based on episomal replication of plasmids. Herein, we present this methodology.

Original languageEnglish
Title of host publicationIn Vitro Mutagenesis Protocols
Subtitle of host publicationThird Edition
EditorsJeff Braman
Pages87-100
Number of pages14
Edition3
ISBN (Electronic)9781607616528
DOIs
Publication statusPublished - 1 Jan 2010

Publication series

NameMethods in Molecular Biology
Volume634
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Attb and attP sites
  • Bxb1 integrase
  • Integrative recombination
  • Malaria
  • Plasmodium falciparum
  • Transfection
  • Transgene expression

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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