Document Detail


Efficient site-specific integration in Plasmodium falciparum chromosomes mediated by mycobacteriophage Bxb1 integrase.
MedLine Citation:
PMID:  16862136     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Here we report an efficient, site-specific system of genetic integration into Plasmodium falciparum malaria parasite chromosomes. This is mediated by mycobacteriophage Bxb1 integrase, which catalyzes recombination between an incoming attP and a chromosomal attB site. We developed P. falciparum lines with the attB site integrated into the glutaredoxin-like cg6 gene. Transfection of these attB(+) lines with a dual-plasmid system, expressing a transgene on an attP-containing plasmid together with a drug resistance gene and the integrase on a separate plasmid, produced recombinant parasites within 2 to 4 weeks that were genetically uniform for single-copy plasmid integration. Integrase-mediated recombination resulted in proper targeting of parasite proteins to intra-erythrocytic compartments, including the apicoplast, a plastid-like organelle. Recombinant attB x attP parasites were genetically stable in the absence of drug and were phenotypically homogeneous. This system can be exploited for rapid genetic integration and complementation analyses at any stage of the P. falciparum life cycle, and it illustrates the utility of Bxb1-based integrative recombination for genetic studies of intracellular eukaryotic organisms.
Authors:
Louis J Nkrumah; Rebecca A Muhle; Pedro A Moura; Pallavi Ghosh; Graham F Hatfull; William R Jacobs; David A Fidock
Related Documents :
15314236 - The rd1 virulence locus of mycobacterium tuberculosis regulates dna transfer in mycobac...
18922896 - Characterization of naturally occurring hpv16 integration sites isolated from cervical ...
7840756 - Most of the avian genome appears available for retroviral dna integration.
10225926 - Green fluorescent protein as a marker in plasmodium berghei transformation.
2037296 - Mapping of human chromosome 22 with a panel of somatic cell hybrids.
1996126 - Recommended protocols based on a survey of current practice in genotoxicity testing lab...
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Nature methods     Volume:  3     ISSN:  1548-7091     ISO Abbreviation:  Nat. Methods     Publication Date:  2006 Aug 
Date Detail:
Created Date:  2006-07-24     Completed Date:  2006-09-05     Revised Date:  2014-09-08    
Medline Journal Info:
Nlm Unique ID:  101215604     Medline TA:  Nat Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  615-21     Citation Subset:  IM    
Data Bank Information
Bank Name/Acc. No.:
GENBANK/DQ813651;  DQ813652;  DQ813653;  DQ813654;  PF070036
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Chromosomes / genetics*
Genetic Engineering / methods*
Integrases / genetics*
Molecular Sequence Data
Mutagenesis, Site-Directed / methods*
Mycobacteriophages / genetics*
Plasmodium falciparum / genetics*
Recombination, Genetic / genetics*
Repressor Proteins / genetics*
Transgenes / genetics
Viral Proteins / genetics*
Grant Support
ID/Acronym/Agency:
P01 AI060342/AI/NIAID NIH HHS; P01 AI060342-040002/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Repressor Proteins; 0/Viral Proteins; 0/gp69 protein, Mycobacteriophage Bxb1; EC 2.7.7.-/Integrases
Comments/Corrections
Comment In:
Trends Microbiol. 2007 Jan;15(1):3-6   [PMID:  17126551 ]
Erratum In:
Nat Methods. 2006 Sep;3(9):763

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  High-throughput screening methodology for the directed evolution of glycosyltransferases.
Next Document:  A comprehensive library of fluorescent transcriptional reporters for Escherichia coli.