RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_0306000; Gene model (P.falciparum): PF3D7_0208900; Gene product: 6-cysteine protein (P230p)
PhenotypeNo phenotype has been described
Last modified: 15 April 2010, 07:17
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 20386715
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
The mutant parasite was generated by
Name PI/ResearcherM.R. van Dijk; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-351
Principal name204cl1; 216 (uncloned)
Alternative nameΔp230p-I
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

The mutant lacks expression of P230p.

Protein (function)
The P230p protein is a member of a small family of proteins, the 6-cysteine (cys) family of (surface) proteins. The proteins are characterized by domains of roughly 120 amino acids in size that contain six positionally conserved cysteines (6-cys). Although some species of Plasmodium (may) contain unique members of the 6-cys family, ten members have been identified that are conserved both in structure as well as in genome organization throughout the genus. Some of the conserved 6-cys proteins are encoded by genes that form paralogous gene-pairs which are closely linked in the genome separated by less then 2 kb of intergenic region. Most members have a GPI anchor and are predicted membrane surface proteins whereas others appear to be secreted and most members are expressed in a discrete stage-specific manner in gametocytes, sporozoites or merozoites (see also 'Additional Information'). Evidence has been presented that P230p is specifically expressed in gametocytes of P. berghei.

Phenotype analyses indicate that that P230p is not essential in the complete life cycle of the parasite. The mutant lacking P230p shows growth and development characteristics that are comparable to wild type parasites both in the vertebrate host and in the mosquito.

Additional information
The p230p gene forms a paralogous gene pair with the gene p230 (PB000403.00.0; PFB0405w), which are closely linked in the genome. In contrast to the non-essential role of P230p, the paralog P230 plays an important role in male fertility and fertilisation (see mutant RMgm-350 which lacks expression of P230).

The p230p locus is commonly used as a locus for integration of transgenes in P. berghei, for example fluorescent proteins.

The mutant described here, Δp230p-I, contains a partial disrupted p230p gene. A fragment of the gene is deleted from the second 6-cys domain (i.e. the first 894 amino acids are still present in the genome). A second mutant (Δp230p-II; RMgm-352) has been generated and the deleted fragment includes part of the first 6-cys domain (i.e. the first 492 amino acids are still present).

Table: P. falciparum gene members of the 6-cys family

Gene P. falciparum Gene P. falciparum
p48/45  PF13_0247 p12p  PFF0620c
p47  PF13_0248 p230p  PFB0400w
p36  PFD0210c p230  PFB0405w
p52  PFD0215c p38  PFE0395c
p12  PFF0615c p41  PFD0240c

Other mutants
RMgm-350: A mutant lacking expression of the paralog P230 (PB000403.00.0; PFB0405w)
RMgm-352: An independent mutant lacking expression of P230p

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0306000
Gene Model P. falciparum ortholog PF3D7_0208900
Gene product6-cysteine protein
Gene product: Alternative nameP230p
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption Of the 6863bp coding region, nucleotides 3135-3194 were disrupted
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1cggggtaccgaaacaatcgaatttctatgc
Additional information primer 1L1.5 (KpnI); Pbs230p F 5'KO
Sequence Primer 2cccaagcttttggcgtcccatctatgc
Additional information primer 2L1.6 (HindIII); Pbs230p R 5'KO
Additional information primer 3L1.3 (EcoRV); Pbs230p F 3'KO
Sequence Primer 4cgcggatcctactgtaataccttttttccc
Additional information primer 4L1.4 (BamHI); Pbs230p R 3'KO
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6