RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
TaggedGene model (rodent): PBANKA_1441400; Gene model (P.falciparum): PF3D7_1226600; Gene product: proliferating cell nuclear antigen 2 (PCNA2)
Name tag: GFP
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst; Sporozoite; Liver stage;
Last modified: 18 March 2019, 15:45
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) : 30865364
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/ResearcherPradhan S, Dhar SK
Name Group/DepartmentSpecial Centre for Molecular Medicine
Name InstituteJawaharlal Nehru University
CityNew Delhi
Name of the mutant parasite
RMgm numberRMgm-4600
Principal namePbPCNA2GFP::hDHFR
Alternative namePbPCNA2‐GFP
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stagePbPCNA2‐GFP expression in all blood stages (diffused expression in whole parasite)
Gametocyte/GametePbPCNA2‐GFP expression in all blood stages (diffused expression in whole parasite)
Fertilization and ookinetePbPCNA2‐GFP expression in gametocytes and ookinetes. In ookinetes the signal was higher in nucleus than cytoplasm
OocystPbPCNA2‐GFP expression in oocysts (diffused expression in whole parasite)
SporozoitePbPCNA2‐GFP expression in sporozoites (mainly in cytoplasm)
Liver stagePbPCNA2‐GFP expression in liver stages (diffused expression in whole parasite)
Additional remarks phenotype

The mutant expresses a C-terminal GFP-tagged version of PCNA2

Protein (function)
Proliferating cell nuclear antigen (PCNA) proteins play a role in DNA replication and DNA repair pathways. The genome of Plasmodium parasites encodes for two PCNA proteins. P. falciparum PCNA1 (PfPCNA1) has been previously implicated in DNA replication as well as DNA repair, whereas role of P. falciparum PCNA2 (PfPCNA2) has been assigned to DNA repair, during the asexual stage of the parasite.

PbPCNA2‐GFP expression in all stages.

Additional information

Other mutants

  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1441400
Gene Model P. falciparum ortholog PF3D7_1226600
Gene productproliferating cell nuclear antigen 2
Gene product: Alternative namePCNA2
Details of the genetic modification
Name of the tagGFP
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 markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationIn order to generate the PCNA2 knockout‐targeting construct, a920 bp fragment was amplified from the 5′UTR region of PbPCNA2 and a 609 bp from the 3′UTR of the gene. Both UTRs were cloned in knockout vector flaking the GFP and DHFR cassette.

For generating the PbPCNA2‐GFP tagging construct, 1898 bp fragment, containing 999 bp of 5′UTR and full length PbPCNA2 coding sequence excluding the stop codon, was amplified from the parasite genomic DNA.
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