Summary

RMgm-1374
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1136700; Gene model (P.falciparum): PF3D7_1360500; Gene product: guanylyl cyclase beta (GCbeta; GCβ)
Transgene
Transgene not Plasmodium: GFP
Promoter: Gene model: PBANKA_0711900; Gene model (P.falciparum): PF3D7_0818900; Gene product: heat shock protein 70 (HSP70)
3'UTR: Gene model: Not available; Gene product: Not available
Insertion locus: Gene model: PBANKA_1136700; Gene product: guanylyl cyclase beta (GCbeta; GCβ)
Phenotype Sporozoite; Liver stage;
Last modified: 5 April 2016, 10:42
  *RMgm-1374
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 26796412
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/ResearcherKooij TWA; Kenthirapalan S
Name Group/DepartmentMolecular Parasitology - Departement of Medical Microbiology
Name InstituteRadboud University Medical Centre
CityNijmegen
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-1374
Principal namegcβ–
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNo sporozoites in vivo
Liver stageNo transmission by bite of infected mosquitoes
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of GCβ and expresses GFP under control of the constitutive hsp70 promoter

Protein (function)
Membrane transport proteins (MTP) transfer compounds across biological membranes and encompass diverse gene families, namely ion channels, ATP-dependent pumps and secondary active porters including those of the major facilitator superfamily. In contrast to bacteria, archaea and fungi, parasitic protozoa such as Trypanosoma brucei and the malaria parasite Plasmodium falciparum allocate only a small proportion of their genomes (2–3%) to membrane transport. P. falciparum encodes at least 122 MTPs. For 39 gene products, functional or subcellular localization predictions remain elusive, rendering them orphan MTPs.
In this study a comprehensive genetic analysis is reported of 35 orphan transport proteins of Plasmodium berghei during its life cycle in mice and Anopheles mosquitoes.

Phenotype:
No sporozoites in vivo; No transmission by bite of infected mosquitoes;

Additional information

Other mutants
See also other mutants: link


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1136700
Gene Model P. falciparum ortholog PF3D7_1360500
Gene productguanylyl cyclase beta
Gene product: Alternative nameGCbeta; GCβ
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(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
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr/yfcu
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 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

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
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/yfcu
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0711900
Gene Model P. falciparum ortholog PF3D7_0818900
Gene productheat shock protein 70
Gene product: Alternative nameHSP70
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
3'-UTR
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative name
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionInsertion locus
Gene Model of Parasite PBANKA_1136700
Gene productguanylyl cyclase beta
Gene product: Alternative nameGCbeta; GCβ
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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