News 2004 - 2006

  •  09/2006

The Leiden Malaria Group and collaborators published in Science

Mair et al. (2006). Regulation of sexual development of Plasmodium by translational repression. Science 313: 667-9

Timing the Sexual Development of Parasites (pdf), Gevoelige achilleshiel (pdf), Klaargelegd voor later (pdf)


  • 09/2006

New insights in regulation of gene expression during sexual development of malaria parasites.

A protein with homology to the DDX6 family of RNA helicases is involved in translational repression of mRNA in female gametocytes and in regulating sexual development (Mair et al, (2006). Regulation of sexual development of Plasmodium by translational repression. Science 313: 667-9).

Timing the Sexual Development of Parasites, Gevoelige achilleshiel (pdf), Klaargelegd voor later (pdf)

  • 09/2006

Several (genetically modified) P. berghei parasite lines has been deposited at MR4

  1. Reference lines of the ANKA strain of  P. berghei (cl15cy1; HPEcy1m50cl1)
    • The cl15cy1 clone is a reference clone of the ANKA strain and used by different labs. It is derived from the 8417HP clone and produces wild type numbers of gametocytes, ookinetes, oocysts, sporozoites. This line has been used to sequence the genome of P. berghei (Hall et al., 2005; Science 307. 82-6). 
    • The HPEcy1m50cl1 (ANKA strain) is also derived from 8417HP and produces no gametocytes. It has been used in our lab as a reference non-gametocyte producer clone and has also been used to analyse transcription in pure, synchronised asexual blood stages (Hall et al., 2005; Science 307. 82-6)
  2. Genetically modified lines/clones
    • Four published, 'reference' lines (259cl2 (=GFPcon); 354cl4; 507m6cl1; 676m1cl1) that express reporter genes GFP or the fusion protein GFP-Luciferase under different promoters (Janse et al., 2006; Mol Biochem Parasitol. 145: 60-70). These have been sent to different labs and are used to visualise parasite development.
    • Two published knock-out parasites lines (137cl8; 270cl1) lacking the expression of the gametocyte/gamete specific proteins P48/45 and P47 respectively (van Dijk et al. 2001, Cell 104, 153-164; Khan et al., 2005, Cell 121: 675-87 ). Since these lines are defective in either male or female fertility, these lines are used by different labs for crossing experiments to test the fertility of gametes of other knock-out mutants.
  • 09/2006

 Two videos showing P. berghei protocols have been placed onto the password protected area of our website

  • Purification of cultured mature schizonts using Nycodenz-gradient centrifugation
  • Transfection of purified schizonts using the Amaxa Nucleofector device

  • 07/2006

Three detailed protocols from our group have been published in Nature Protocols.

High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium berghei

Selection by flow-sorting of genetically transformed, GFP-expressing blood stages of the rodent malaria parasite, Plasmodium berghei

Real-time in vivo imaging of transgenic bioluminescent blood stages of rodent malaria parasites in mice


  • 07/2006

A negative selection marker has been developed in the Leiden Malaria Group for the use in reverse genetics of Plasmodium berghei

yFCU; which combines a fusion of yeast cytosine deaminase and uridyl phosphoribosyl transferase (UPRT) with in vivo selection using the prodrug 5-fluorocytosine (5-FC). The combination of a positive and negative selectable marker allows for rapid analysis of the phenotype by targeted disruption of a gene and further associate phenotype and function by genotype restoration (Braks JA et al., Nucleic Acids Res. 34: e39.) This negative selection system is also adapted to facilitate drug-selectable marker (vector) recycling which in principle will allow unlimited manipulation of a single parasite clone. A standard vector for genetic modification of P. berghei has been made that contains a fusion of the negative selectable marker yFCU with the positive selectable marker hDHFR and which allows for excision of the drug-selectable markers from the genome through homologous recombination. In this manner it should also prove possible to perform ‘hit and run’ mutagenesis on P. berghei. This vector has been send to MR4 and is described in the plasmid database on our Sharpoint environment.


  • 07/2006

The plasmid database on the password protected area of our website has been updated

and all plasmids have been sent to MR4. It also contains a standard vector for drug-selectable marker recycling using positive/negative selection


  • 03/2006

Information about the organization of the genomes of the rodent malaria parasites (RMP) genomes is placed onto the password protected area of our website

This information consists of:

  1. The chromosome location of all predicted P. berghei, P. yoelii and P. chabaudi genes with P. falciparum orthologs. The position of these genes is described relative to the chromosomal location of their P. falciparum orthologs (2 Excel files).
  2. All available RMP sequence contigs with sequence homology to P. falciparum aligned along the composite chromosomes of rodent malaria parasite (RMP) and aligned along the P. falciparum chromosomes (information in 2 Excel files)
  3. A schematic representation of the synteny map of the RMP genome and the P. falciparum genome, showing all 14 chromosomes, regions of synteny, breakpoints of synteny, centromere locations etc (pdf file).
  4. Lists of P. falciparum specific genes for which an ortholog has not been found in the RMP genome and also a list of RMP-specific intersyntenic genes (information in 4 tables; P. falciparum subtelomeric genes, intersyntenic genes and intrasyntenic genes; RMP intersyntenic genes)

All information is based on the published study by Kooij et al (2005). A Plasmodium whole-genome synteny map: Indels and synteny breakpoints as foci for species-specific genes; PLoS Pathogens, 23, 1(4):e44


  • 02/2006

PhD thesis: Taco Kooij (LUMC, Leiden). Rodent Malaria Parasites: Genome Organization and Comparative Genomics.

Genome Organization & Comparative (pdf)

  • 01/2006

The Leiden Malaria Group and collaborators published in PLoS Pathogens

The Leiden Malaria Group and collaboratorsKooij et al (2005). A Plasmodium whole-genome synteny map: Indels and synteny breakpoints as foci for species-specific genes. PLoS Pathogens, 23, 1(4):e44published in PLoS Pathogens


  • 11/2005

All standard plasmids for genetic modification of P. berghei from the Leiden Malaria group are now available from the Malaria Research and Reference Reagent Resource Center (MR4)

Detailed information about these plasmids can be found in an excel file on the password protected area of our website.


  • 11/2005

Several novel technologies have been published that improve larger scale genetic modification of P. berghei

  1. Sakamoto H. et al. (2005). Towards systematic identification of Plasmodium essential genes by transposon shuttle mutagenesis. Nucleic Acids Res., 33(20):e174. (from the group of Robert Menard, Pasteur Institute, Paris).
  2. Ecker A. et al. (2005). Generation of gene targeting constructs for P. berghei by a PCR-based method amenable to high throughput applications. Mol Biochem Parasitol. 145(2): 265-8 (from the group of Oliver Billker and Bob Sinden, Imperial College, London).
  3. Janse C. J. et al.. (2005). High efficiency transfection of P. berghei facilitates novel selection procedures. Mol Biochem Parasitol. 145(1): 60-70 (from the group of Andy Waters and Chris Janse, Leiden University Medical Center, Leiden)

  • 08/2005

The Leiden Malaria Group and collaborators published in PNAS

van Dijk, M.R. et al (2005). Genetically attenuated, P36p-deficient malarial sporozoites induce protective immunity and apoptosis of infected liver cells. Proc Natl Acad Sci U S A (102:12194-9)

Malaria Vaccines: Back to the Future? (pdf), Herkansing voor malariavaccin (pdf)


  • 08/2005

The Leiden Malaria Group used Plasmodium berghei is used as a model to investigate genetically modified, attenuated sporozoites as a vaccine

van Dijk, M.R. et al (2005). Genetically attenuated, P36p-deficient malarial sporozoites induce protective immunity and apoptosis of infected liver cells. Proc Natl Acad Sci U S A (102:12194-9)

Malaria Vaccines: Back to the Future? (pdf), Herkansing voor malariavaccin (pdf)


  • 08/2005

Information on standard plasmids for genetic modification of P. berghei on our website is updated

The excel database contains information of plasmids (sequence, plasmid maps etc.) used for disruption, mutating or tagging of genes and for expression of transgenes, including tags such as TAP, c-myc and GFP and transgenes such as GFP, RFP and luciferase.

read more (password protected area)


  • 08/2005

The Leiden Malaria Group and collaborators published in PNAS

Franke-Fayard B.  et al. (2005). Murine malaria parasite sequestration: CD36 is the major receptor, but cerebral pathology is unlinked to sequestration. Proc Natl Acad Sci U S A. 102. 11468-73

Deep-organ malaria parasite not associated with cerebral complications (pdf)


  • 08/2005

The Leiden Malaria Group developed a method for in vivo visualization of sequestration of schizonts of P. berghei (sequestered infected blood cells) in life animals using luciferase-expressing parasites

Franke-Fayard B.  et al. (2005). Murine malaria parasite sequestration: CD36 is the major receptor, but cerebral pathology is unlinked to sequestration. Proc Natl Acad Sci U S A. 102. 11468-73

Deep-organ malaria parasite not associated with cerebral complications (pdf)


  • 06/2005

Proteome data Plasmodium berghei male and female gametocytes is placed on our website

Proteome data is available in excel databases with P. berghei gene models linked to their P. falciparum gene orthologs. In addition we present a comparison of proteome data of P. berghei and P. falciparum of different published studies.

read more (password protected area)


  • 06/2005

Phenotype data pertaining to kinase deficient mutant P. berghei parasites

(i.e. either MAP2 or NEK4) has been incorporated into the Excel database which conatins information on all genetically modified P. berghei mutant parasite lines

read more (password protected area)


  • 06/2005

Proteome analysis of separated male and female gametocytes (the precursor cells of the male and female gametes) is published

Shahid M. Khan, Blandine Franke-Fayard, Gunnar R. Mair, Edwin Lasonder, Chris J. Janse, Matthias Mann & Andrew P. Waters. Proteome analysis of separated male and female gametocytes reveals novel sex specific Plasmodium. Cell (2005) 121:675-687

Revealing the Molecular Determinants of Gender in Malaria Parasites (pdf), De malariacirkel (pdf), Proteomes of Plasmodium gametocytes (pdf)


  • 06/2005

The Leiden Malaria Group and collaborators published in Cell

Proteome analysis of separated male and female gametocytes reveals novel sex specific Plasmodium.
Cell (2005) 121:675-687

Revealing the Molecular Determinants of Gender in Malaria Parasites (pdf), De malariacirkel (pdf), Proteomes of Plasmodium gametocytes (pdf)


  • 02/2005

Plasmodium berghei is used as a model to investigate genetically modified, attenuated sporozoites as a vaccine

Mueller AK, Labaied M, Kappe SH, Matuschewski K. Genetically modified Plasmodium parasites as a protective experimental malaria vaccine. Nature (2005)433:164-7.
See also Parasitology. Malaria vaccines: back to the future? (pdf) Waters AP, Mota MM, van Dijk MR, Janse CJ. Science. (2005), 307:528-30.


  • 01/2005

Transcriptome data of the blood stages of Plasmodium berghei (including gametocytes) is placed on our website

An Excel database with P. berghei gene models linked to their P. falciparum gene orthologs

read more (password protected area)


  • 01/2005

The genome of Plasmodium berghei is published

with transcriptome and proteome analyses
Hall N, Karras M, Raine JD, Carlton JM, Kooij TW, Berriman M, Florens L, Janssen CS, Pain A, Christophides GK, James K, Rutherford K, Harris B, Harris D, Churcher C, Quail MA, Ormond D, Doggett J, Trueman HE, Mendoza J, Bidwell SL, Rajandream MA, Carucci DJ, Yates JR 3rd, Kafatos FC, Janse CJ, Barrell B, Turner CM, Waters AP, Sinden RE.
A comprehensive survey of the Plasmodium life cycle by genomic, transcriptomic, and proteomic analyses.
Science (2005) 307:82-6.

Revealing the Molecular Determinants of Gender in Malaria Parasites (pdf), Malaria of mice and men; Using models to understand disease (pdf), Malaria-onderzoekers leren hun vijand kennen (pdf), De familie malaria (pdf)


The Leiden Malaria Group and collaborators published in Science

Hall N, Karras M, Raine JD, Carlton JM, Kooij TW, Berriman M, Florens L, Janssen CS, Pain A, Christophides GK, James K, Rutherford K, Harris B, Harris D, Churcher C, Quail MA, Ormond D, Doggett J, Trueman HE, Mendoza J, Bidwell SL, Rajandream MA, Carucci DJ, Yates JR 3rd, Kafatos FC, Janse CJ, Barrell B, Turner CM, Waters AP, Sinden RE.
A comprehensive survey of the Plasmodium life cycle by genomic, transcriptomic, and proteomic analyses.
Science (2005) 307:82-6

Revealing the Molecular Determinants of Gender in Malaria Parasites (pdf), Malaria of mice and men; Using models to understand disease (pdf), Malaria-onderzoekers leren hun vijand kennen (pdf), De familie malaria (pdf)


  • 01/2005

A picture of the natural host of P. berghei, Grammomys surdaster, on our website

The picture is obtained from the Institute of Tropical Medicine, Department of Parasitology, Antwerp. The animal shown is from a small breeding colony in the Institute of Tropical Medicine in Antwerp that originated from about fifty animals that were trapped in an area of 5 km around Kaindu, 20 km nort west of Mumbwa near the Kafua National Park (Zambia) in 1996. Determination of the species was based on analysis of the cytochrome B sequence of mitochondrial DNA at the Department of Animal Ecology of the University of Antwerp. In some handbooks Grammomys surdaster  is called Grammomys dolichurus (woodland thicket rat) (Kingdon, J. (1974). East African Mammals. Vol.II part B (Hares and Rodents). London, Academic Press; R.M. Nowak, Walker's mammals of the World, vol. 2; Johns Hopkins University Press) but the taxonomy of thicket rats is still in a state of flux and in review. Based on differences in mitochondrial DNA G. surdaster and G. dolichurus are well separated species and the differences warrant to group them in different taxa (pers. comm. prof. dr. H. Leirs, University of Antwerp).
More information on Grammomys dolichurus


  • 11/2004

Improvement of transfection efficiency in P. berghei

Using a new method of transfection, the AMAXA non-viral nucleofectortm technology, we obtain transfection efficiencies in the range of 10-2-10-3 (with the standard transfection technology the efficiency was 10-6-10-9). The methods have not been published yet but are described in our transfection protocols.

read more (password protected area)


  •  11/2004

New on our website

Molecular protocols used in our laboratory are placed here (password protected area)


  • 11/2004

The malariagroup of the LUMC has a new professor, Andy Waters

Andy Waters was promoted to the position of full professor with effect from October 1th 2004.

Malariahoogleraar (pdf)

  •  9/2004

A reference line of P. berghei, expressing Green Fluorescent Protein (GFP) in all life cycle stages

We generated a transgenic line of P. berghei that expresses GFP at a high level in all life cycle stages. This line shows similar growth characteristics to wild type P. berghei parasites of the ANKA strain, both in the vertebrate and mosquito host.
More information: Molecular & Biochemical Parasitology paper (abstract) (pdf); information in our database 'Mutant lines (knockout, transgene, tagging) and natural lines of P. berghei'. article in Cicero: Parasiet geeft licht (pdf)


  • 7/2004

Our website has been updated

 Our website has been updated, in particular we have significantly changed the part on the 'Plasmodium berghei Research Model of Malaria'. A part of the information provided is no longer freely accessible, such as protocols and databases. For this information a password is needed.On a regular basis the password will change and you will be required to register again. It is a simple and brief process but it allows us to keep only active researchers as subscribers. In the password protected part of the website we now provide the following information:
- Detailed protocols on 'P. berghei: general parasitological methods', 'Molecular Methods', 'P. berghei: transfection methods', including information about transfection using the Amaxa method
- Excel databases with information on the basic plasmids we use for transfection of P. berghei and information on all mutant lines (gene knockout, transgene, tagging) that have been made in our laboratory or that have been described in the literature, with emphasis on the phenotype characteristics of the mutant lines
This part of our website is still under construction and comments or additional information are welcome. We will regularly update the protocols and databases and in the future more data concerning genome structure, transcriptomes and proteomes of P. berghei will be added. The relevant data will also be linked in to public databases such as PlasmoDB and GeneDB
Please follow the link and fill in the form for request of the password.

  • 5/2004

Researchers from the Leiden malaria Group contributed to two papers published in Cell

Blandin S, Shiao SH, Moita LF, Janse CJ, Waters AP, Kafatos FC, Levashina EA.
Complement-like protein TEP1 is a determinant of vectorial capacity in the malaria vector Anopheles gambiae. Cell 2004 Mar 5;116(5):661-70.

Billker O, Dechamps S, Tewari R, Wenig G, Franke-Fayard B, Brinkmann V.
Calcium and a calcium-dependent protein kinase regulate gamete formation and mosquito transmission in a malaria parasite. Cell 2004 14;117(4):503-14


  • 4/2007

New book 'Malaria Parasites: Genomes and Molecular Biology' (2004), edited by Andy Waters and Chris Janse

Internationally renown experts provide up-to-date reviews of the most important aspects of post-genomic malaria research: Genomes and Molecular Biology (pdf)
See also review: Postgenetic Plasmodium (pdf)


  • 2/2004

Prize for a poster from Shahid Khan (LUMC, Leiden)

'Sensitive comparative proteomic analysis of highly purified Plasmodium male and female gametocytes' presented at the Molecular Approaches to Malaria' meeting (2004; Lorne, Victoria, Australia) 

Sensitive comparative proteomic analysis of highly purified Plasmodium male and female gametocytes (pdf)