RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
TaggedGene model (rodent): PBANKA_0612400; Gene model (P.falciparum): PF3D7_0411000; Gene product: conserved Plasmodium protein, unknown function (AP2R-1, AP2-R1 (AP2 Transcription Factor-related gene 1), AP2-Z (AP2-zygote))
Name tag: mNeonGreen
Phenotype Gametocyte/Gamete;
Last modified: 26 May 2022, 12:57
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 34119684
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherYuda M, Nishi T
Name Group/DepartmentDepartment of Medical Zoology
Name InstituteMie University School of Medicine
CityMie, Tsu
Name of the mutant parasite
RMgm numberRMgm-5034
Principal nameAP2R-1-mNeonGreen
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot different from wild type
Gametocyte/GameteFluorescence microscopy using transgenic parasites expressing AP2R-1 as a mNeonGreen-tagged protein showed that it is expressed in the nucleus of female gametocytes.
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

The mutant expresses a C-terminal mNeonGreen-tagged version of AP2R-1

See also mutants RMgm-5199 and RMgm-5200 targeting this gene by the same group with a different expression pattern when the gene was tagged with GFP (as a result of translational repression in female gametocytes).

Protein (function)
Through a screen, a gene containing a putative AP2 domain was identified (PBANKA_0612400). Although the E- value obtained from SMART analysis was not small enough to conclude that it is an AP2 domain (E-value =0.042),  the domain has a structural characteristics of AP2 domain: three antiparallel β-sheets followed by a single α-helix.  A unique feature in this putative AP2 domain was a  long insertion sequence between predicted sheets 2 and 3.  Assuming that the high E-value is attributed to this sequence, we deleted it from the original sequence of the putative AP2 domain  and subjected the modified sequence to SMART analysis. As a result, a much smaller E-value  (E-value <1 ×105) was obtained from the putative AP2 domain, suggesting that this gene, hereafter referred to as AP2 TF-related gene 1 (AP2R-1), is a novel member of the Plasmodium AP2 family.


Fluorescence microscopy using transgenic parasites expressing AP2R-1 as a mNeonGreen-tagged protein showed that it is expressed in the nucleus of female gametocytes.

Additional information

From the Abstract:

'Fluorescence microscopy showed that AP2-G expression was first observed in the parasite 12 h after erythrocyte invasion and peaked at 18 h when sexual features were first manifested in  early gametocytes. Expression of  AP2-G decreased with manifestation of  sex-specific features. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) was performed at peak AP2-G expression and identified over 1000 binding sites in the genome. The main binding motif of the TF predicted from the binding sites was GTACNY. Predicted targets contained a number of genes related to protein biogenesis, suggesting that AP2-G plays a role in establishing a cellular basis required for sexual differentiation. AP2-G binding sites also existed upstream of gametocyte-specific TFs, namely AP2-G2, AP2-FG, and AP2-G itself. Furthermore, the target contained two AP2 TF-related genes. Disruption of  these genes resulted in  the arrest of  ookinete development. These results suggest another role of  AP2-G: activating a  transcriptional cascade to  promote conversion into early gametocytes.'

Other mutants

  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0612400
Gene Model P. falciparum ortholog PF3D7_0411000
Gene productconserved Plasmodium protein, unknown function
Gene product: Alternative nameAP2R-1, AP2-R1 (AP2 Transcription Factor-related gene 1), AP2-Z (AP2-zygote)
Details of the genetic modification
Name of the tagmNeonGreen
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markerunknown
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 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6