RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-36
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1400800; Gene model (P.falciparum): PF3D7_1302200; Gene product: early transcribed membrane protein 13 (UIS3; up-regulated in infective sporozoites; ETRAMP13)
Phenotype Liver stage;
Last modified: 19 February 2009, 20:37
  *RMgm-36
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 15580261
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei NK65
Name parent line/clone Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherA.K. Mueller, S.H.I. Kappe, K. Matuschewski
Name Group/DepartmentDepartment of Parasitology
Name InstituteHeidelberg University School of Medicine
CityHeidelberg
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-36
Principal nameuis3-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageIn vitro invasion of hepatocytes by the sporozoites is not affected. Liver stage development is strongly impaired. Sporozoites initiate the transformation of the banana-shaped, elongated sporozoite to a spherical liver trophozoite. However, they show a severe defect in their ability to complete transformation into liver trophozoites. They lack the capacity to develop into mature liver schizonts.
Infection of highly susceptible, young Sprague-Dawley rats by intravenous inoculation of high numbers of sporozoites (100.000) did not result in blood stage infection.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of the UIS3 protein ('up-regulated in infective sporozoites'; early transcribed membrane protein 13, ETRAMP13).

Protein (function)
UIS3 has been identified by transcription-profiling of sporozoites (Kaiser et al., (2001). Mol. Microbiol. 51, 1221-32).

Phenotype
The phenotype analysis demonstates a role of this protein in liver stage development.

Additional information
A P. yoelli mutant has been generated that lacks the expression of this protein (RMgm-37). This mutant has the same phenotype as described above.
P. berghei and P. yoelii 'attenuated sporozoites' lacking expression of UIS3 can induce sterile (protective) immunity in mice against challenge with wild type sporozoites by immunization of mice (C57Bl/6; BALB/c) with the mutant sporozoites.

Other mutants
A P. yoelli mutant has been generated that lacks the expression of this protein (RMgm-37).
A. P. berghei mutant has been generated that lacks the expression of not only UIS3 but also UIS4 (RMgm-39).

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1400800
Gene Model P. falciparum ortholog PF3D7_1302200
Gene productearly transcribed membrane protein 13
Gene product: Alternative nameUIS3; up-regulated in infective sporozoites; ETRAMP13
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 parasitetgdhfr
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 15'-GGGTACCCGCATTAGCATAACATCTCATTGG-3'
Additional information primer 1UIS3rep1for
Sequence Primer 25' -
CAAGCTTGCTTTCATATATTTGTTATTTGTC-3'
Additional information primer 2UIS3rep2rev
Sequence Primer 35'-GGAATTCCCATATGTTTGTGTAACATC-3'
Additional information primer 3UIS3rep3
Sequence Primer 45' -CTCTAGAGTGTGCTTAAATGTTTCTTTAAAC-3'
Additional information primer 4UIS3rep4rev
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren