RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1343
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene not Plasmodium: nahG
Promoter: Gene model: Not available; Gene model (P.falciparum): PF3D7_0709000; Gene product: chloroquine resistance transporter (CRT)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Replacement locus: Gene model: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Phenotype Asexual bloodstage;
Last modified: 25 October 2015, 13:44
  *RMgm-1343
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 26466097
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 Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherMatsubara R; Nagamune K
Name Group/DepartmentDepartment of Parasitology
Name InstituteNational Institute of Infectious Diseases, Shinjyuku
CityTokyo
CountryJapan
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Name of the mutant parasite
RMgm numberRMgm-1343
Principal namenahG-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageEvidence is presented for increased virulence in mice
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses the nahG gene, a gene that encodes a salicylic acid-degrading enzyme of the plant pathohgenic bacteria Pseudomomas putida (PMID: RMgm-17757215)

Protein (function)
The mutant expresses the nahG gene, a gene that encodes a salicylic acid-degrading enzyme of the plant pathohgenic bacteria Pseudomomas putida (PMID: RMgm-17757215)

Phenotype
Evidence is presented for increased virulence of blood stages.

Additional information
(Some) evidence is presented that P. berghei wild type blood stages produce salicylic acid (SA). No comparison is provided on SA levels in wild type and mutant parasites .

Other mutants

  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene namenahG
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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
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification500 bp sequences corresponding to the 5′ and 3′ regions of the Pb230p gene were amplified by PCR and inserted into the pL0006 vector. Promoter of P. falciparum chloroquine-resistant transporter gene (pCRT), terminator of P. berghei DHFR gene (PbDT), and nahG-HA were amplified and joined by PCR with overlapping primers. The amplicon was also ligated into pL0006. For the control experiment, gfp-HA was amplified and introduced as for nahG. The constructed vector was cut by SacII and NotI, and electroporated into P. berghei ANKA. Transfectants were selected by pyrimethamine. Expression was confirmed by Western blotting with anti-HA antibody.
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite Not available
Gene Model P. falciparum ortholog PF3D7_0709000
Gene productchloroquine resistance transporter
Gene product: Alternative nameCRT
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
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3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
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 / InsertionReplacement locus
Gene Model of Parasite PBANKA_0306000
Gene product6-cysteine protein
Gene product: Alternative name230p
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
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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