RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-236
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0417100; Gene model (P.falciparum): PF3D7_0904300; Gene product: conserved protein, unknown function (CRAGS, CRMP-related antigen of gametocytes and schizonts)
PhenotypeNo phenotype has been described
Last modified: 30 August 2009, 16:36
  *RMgm-236
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Not published (yet)
MR4 number
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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).
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The mutant parasite was generated by
Name PI/ResearcherC.J. Janse, J. Thompson
Name Group/DepartmentInstitute of Immunology and Infection Research
Name InstituteUniversity of Edinburgh
CityEdinburgh
CountryUnited Kingdom
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Name of the mutant parasite
RMgm numberRMgm-236
Principal namePbCRAGSko (1&2)
Alternative name451, 512, 633
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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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 stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of CRAGS (CRMP-related antigen of gametocytes and schizonts; cysteine repeat modular protein, putative).

Protein (function)
CRAGS is related to the Cysteine Repeat Modular Proteins (CRMPs; Thompson et al. (2007) Cell Microbiol. Jun;9(6):1466-80). See also mutants lacking CRMP1 (RMgm-198) and CRMP2 (RMgm-199).

Phenotype
The phenotype analyses indicate that CRAGS is not essential for completion of the Plasmodium life cycle. The phenotype of the mutants has not been analysed in detail and the mutans are not cloned.

Additional information
Two gene models: PB000791.02.0 + PB000813.03.0

See 'Additional Information genetic modification' for the sequence of the correctly spliced protein (verified by RT-PCR of P. berghei ANKA schizont and gametocyte RNA).

Mutants have been generated with a GFP- or TAP-tagged crags gene (lines 510, 511). Both the genotype and phenotype of these mutants have not been analysed in detail.

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0417100
Gene Model P. falciparum ortholog PF3D7_0904300
Gene productconserved protein, unknown function
Gene product: Alternative nameCRAGS, CRMP-related antigen of gametocytes and schizonts
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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 modificationThe correctly spliced protein (verified by RT-PCR of P. berghei ANKA schizont and gametocyte RNA) is: MILLTVLIYFLFRQTKIVILKKMKIYFWNKIKMLALIIHLFVNRNKYLDILLTIDYKERT
TRILNGKCRAGERLTYDYKCESCKKGFYNFSRENNKCFPCPLGTFNDKLGSVMCVNC
PHGYTTHYTGSKSIAECVCDVGFKFDKITNMCLQCNPLEFCDDNERVWSFKNMCMKKKAKEYVKLCEETEKKEIYNLN
IFCPKKGVCLNMEKNKSCVDGNKLIQCKICEENYRYDLISPAINSCVYCNWASYLAIFYFMTIMILINSLRVIYINNL
EITRIINAFIKYIHYISLLRYINSNYDNGIINVVYLFTVGIPLNEILDCIFTGGTNSERIVQKVNFLLCCPLLFSALT
ILGTSIVFLIRKNKKDSKNGEKRLMINPCHFIDKKLAKKTSTDIKFSSPFNMNIDNFEMNNINVEMTIDEDEQYDIDN
LKKCFILFYILYNNFLYFEIIRLCIFFGFCNYDENRKEYFLIIDDSLKCSEIGYYGKVIIIIVFCTCIKFMNYFFVLF
KNNKNTWIVKDILLLYQKIEKFNKVDNILLLFFLILFYKRFLNNSNPSLIYNYKNGLYNGNIHIFQYILIIILFFIYI
LIMYLYVYEFNDSVCGKYENSCNSEKLNSEKLAVKDKKKQYKIIKCISNNLHSCNNLTIDNVEIDKNEKGSGKNNYIF
NAIIPEEDKKEPKNDSKAISNNWNKTDKLNNNNDIGMIKMKIFLSKLIQKVKNPTMNKIESNDKKLDQFLFSFSFFLY
GTILLSYYLFLSYIHINKFAGILTIIVIACNVAIYVFLFSKFIIYFKDDIKKKCERALNIIIKQIRRPGLIGIGTSQK
NTHIDIFKDVKKSDNSLKEKKTKMEKDSVLNKENKNKKSINNIIKKNLGYNYYKQNFKGINSFVENNHKNIKNKAKRY
KLLNISKVWIYFLYNIIDLSNEREMCLAMLIFLTLQNEFFNMNLDRCLEALENNKFYKISNLKLVLKNFIKYRNKFLK
RLSENSNMSIRFNNCEVGTKFDEIKSNNNMNNNIICEDYENLIVFAWGMSILCYCNLKDINYNLSNILLYVSNVLNTC
RYFKNIVDIYFGVYYNYEFSHANKNSIEHNVKWNLYPYNNLDFNNSYNYIIPNSNLNVMDKENNNIQIYSKDFQEPFY
NDMGKSVYSPNEEMIFTKNNRMLTSVDIRGDVMDYTTKGEYNNKGSFSGLLINENMKNIEHRFVRNQNKQLCRIYIHC
LLVDIGELYFNIHYNYTCEKYICNNLLNLWGYNISNNLYMKRYFENIDKFNKNNMMDILLYNNDININIDSNFVLNIS
DYINKNVINIDFYNIFKKIGKRHRIQIKKLIKEGDYLNYNSLVEFSKIYTDYDEIEKIKNIEVDNKIKNDFFKVFSNA
FPIFIEKDNYKYLDMNEISKIEEKKKFIILYKTMGEKYIEEYILKNYEGSKINYFQMDSYGAVISICGSFFKNLLKIE
ENKFEKMENNTDEDMLKKEMVKRERYLVNNNITENDLLNKIHLKMINKNEYNYTNCEFLENKRIGIFSHNNYECNEGV
TGDLKGIDLRNGNSLIGIFPSVILPNAFTIELWLYFNSKKKNNNNLKSFIVCDKNGNSIFVIDRKNDKIESIGVFSNE
ISFLSENYKRRYYICNENNKCEKKEKKNLVINQKVYNGIYIQSEKLKKWEKINKIVKINKWNLINVTKCATGLVYYIN
GKYLSNISHEILNLEKSFEICIFGNSCFGNNNVGLFSSFKIFEFLNNEEIKKRYKLIKSVKNKEMIGDNINMEKNNNV
IEGIDIKMNYMKGNDEEYFIYFIQSQKNKYKVVPTINNPKYWIKIYQLKFVIIKNECNKMKENRNKENIRMKYHFKII
NIDNMNEYGFDFYYKKIGKKLDGVPKIYGINIFSDYLIFGCDLSQFYTIVLYPSLCIYNELVKPCGYTISGWVFFPVE
KSISFLSLISGTNDVHICVFSDDMILRSIENYVRSNKNNERCTVYHSSGFSIKDIKKGWYYLSVVGTLNGQFYFINGC
FKGYHKFCSFDNIKYIGNTSLFINPFPYICFIKVIGRVLSVNEILHEYSISSGYNNFTYLYYYYYLFILFSNNEENDD
ESFVSKNILIDRSHTRTDTISSDDTYSFYCYNKEKYIFFYITKNYNVYIYPLEESKNYYFTLSLVSMINKKLYIFNSI
NDQINNLNIYFINYIVLPEQWTIFTLINIPYINEANYHCLVGGINGRSHIAIDNDDMTLGVLENEEVFKNKNIYEQNE
KYNNDIKGDDYGNFTQIRYKEAYQKFYSCGYKFENPLNKNILLTTRCINNQQTFFINNTNVGTCRSCRSAITCFGNCL
STNNEYSAPFGNYKFIKIVFEYINDDKIKEFYTNYSL
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 1AATAGTAAGCATAAATCTGG
Additional information primer 1
Sequence Primer 2TACATAAGTAAGTCTTTCTCCCG
Additional information primer 2
Sequence Primer 3TAGTATTACCTGAACAATGG
Additional information primer 3
Sequence Primer 4TTTGTAATTACCAAAAGGAGC
Additional information primer 4
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