RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
TaggedGene model (rodent): PBANKA_1104100; Gene model (P.falciparum): PF3D7_0504500; Gene product: MOLO1 domain-containing protein, putative (TPM2)
Name tag: GFP
Phenotype Fertilization and ookinete;
Last modified: 1 September 2020, 15:22
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) : 32736136
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 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
The mutant parasite was generated by
Name PI/ResearcherTremp AZ, Dessens JT
Name Group/DepartmentDepartment of Infection Biology, Faculty of Infectious and Tropical Diseases
Name InstituteLondon School of Hygiene & Tropical Medicine
Name of the mutant parasite
RMgm numberRMgm-4865
Principal nameTPM2-GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteTMP2-GFP located in the crystalloid of ookinetes
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

The mutant expresses a C-terminal GFP-tagged version of TPM2

Protein (function)
The protein TPM2 was identified in a GFP-affinity pulldown with high accuracy mass spectrometry using the GFP-tagged ookinete crystalloid protein LAP3 (PBANKA_0204500).

This protein possesses a central TPM domain named after its founding proteins TLP18.3, Psb32 and MOLO-1 (Pfam 04536), as well as a transmembrane helix near its carboxy terminus.

TPM2-GFP located in the crystalloid of ookinetes

Additional information

Other mutants

  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1104100
Gene Model P. falciparum ortholog PF3D7_0504500
Gene productMOLO1 domain-containing protein, putative
Gene product: Alternative nameTPM2
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 single 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 modificationThe entire coding sequence of PBANKA_1104100 plus ca. 0.6 kb of upstream sequence was PCR amplified from genomic DNA with primers pDNR-110410-F (ACGAAGTTATCAGTCGAGGTACCGCTCAAACTATTCCTCCTCAATC) and pDNR-110410-R (ATGAGGGCCCCTAAGCTGTTTATTCTATATACAACAGTGATTAAATATACAATG) and cloned into SalI/HindIII-digested pDNR-EGFP by in-fusion cloning to give plasmid pDNR-TPM2/GFP. The 3'UTR of of this gene was amplified with primers pLP-110410-F (ATATGCTAGAGCGGCCATGTATGATATGTATTTTTTGCG) and pLP-110410-R (CACCGCGGTGGCGGCCAATTAAATGAAACTGCGGCAC) and the resulting fragment cloned into NotI-digested pLP-hDHFR by in-fusion cloning to give plasmid pLP-hDHFR/TPM2.
The tpm2-specific sequence from pDNR-TPM2/GFP was transferred to pLP-hDHFR/TPM2 by Cre/loxP recombination to give the final construct pLP-TPM2/GFP. This plasmid was digested with KpnI and SacII before gene targeting by double crossover homologous recombination.
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