RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
Genetic modification not successful
TaggedGene model (rodent): PBANKA_0708100; Gene model (P.falciparum): PF3D7_0822500; Gene product: dynein light chain 1 | leucine-rich repeat protein (PbDLC1; DLC1; PfLRR7)
Name tag: c-myc
PhenotypeNo phenotype has been described
Last modified: 11 May 2010, 09:19
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene tagging
Number of attempts to introduce the genetic modification 2
Reference (PubMed-PMID number) Reference 1 (PMID number) : 20421304
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 676m1cl1 (RMgm-29)
Other information parent lineP.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter (PubMed: PMID: 16242190).
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherW. Daher; J. Khalife
Name Group/DepartmentUnité INSERM 547 and Center for Infection and Immunity of Lille INSERM U1019 - CNRS UMR 8204
Name InstituteUniv Lille Nord de France, Institut Pasteur de Lille

  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0708100
Gene Model P. falciparum ortholog PF3D7_0822500
Gene productdynein light chain 1 | leucine-rich repeat protein
Gene product: Alternative namePbDLC1; DLC1; PfLRR7
Details of the genetic modification
Name of the tagc-myc
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/constructPlasmid 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 markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationNegative attempts to tag the dlc1 gene of P. berghei with GFP suggest that dlc1 is essential for blood stage development. In the same study negative attempts are reported to disrupt or tag the dcl1 gene of P. falciparum.

Dynein light chain 1 (LC1), a member of the Leucine Rich Repeat protein family, has been shown to be engaged in controlling flagellar motility in Chlamydomonas reinhardtii and Trypanosoma brucei via its interaction with the dynein-gamma heavy chain.
By screening for P. falciparum genes expressing Leucine Rich Repeat proteins a candidate dynein LC1 ortholog was identified (PfLRR7; Daher et al., 2007, Mol. Biochem. Parasitol. 155, 161-6). DLC1 of P. falciparum and P. berghei is expressed in gametocytes. In P. berghei DCL1 is specifically expressed in male gametocytes, indicating a role in flagellar motility in male gametes.
Negative attempts to tag or disrupt the dlc1 gene by reverse genetic approaches in both P. falciparum and P. berghei suggest either an essential function for blood stage development or the inaccessibility of its locus. Expression studies revealed high levels of DLC1 protein in late trophozoites and schizonts, pointing to an unexpected role of this protein in blood-stage parasites as they do not have flagella. Interactions studies and co-immunoprecipitation experiments revealed that PfDLC1 was able to bind to P. falciparum myosin A and actin 1. These observations suggest an unexpected role of PfDLC1 in the
motor complex involving actin/myosinA during blood stage development.
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
Additional information primer 1Pb1(F)(KpnI); c-terminal part of PbDLC1
Additional information primer 2Pb2(R)(ApaI); c-terminal part of PbDLC1
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