RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1327

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_0931200; Gene model (P.falciparum): PF3D7_1116800; Gene product: heat shock protein 101 \| chaperone protein ClpB2 (HSP101) Name tag: mCherry-3xMyc
Phenotype	Asexual bloodstage;

Last modified: 5 April 2016, 09:33

*RMgm-1327

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 26219962
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	P. berghei ANKA cl15cy1
Other information parent line	A 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/Researcher	Matz, JM; Kooij, TW
Name Group/Department	Medical Microbiology
Name Institute	Radboudumc
City	Nijmegen
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-1327
Principal name	hsp101-mCh(sens)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Abundant expression at the PVM in younger blood stages and in motile tubular structures in maturing trophozoites
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a mCherry tagged version of HSP101 (and lacks the hdfr-yfcu drug-selectable marker that has been removed by negative selection) Protein (function) Plasmodium parasites remodel their vertebrate host cells by translocating hundreds of proteins across an encasing membrane into the host cell cytosol via a putative export machinery termed PTEX (Plasmodium Translocon of EXported protein). HSP101 (PbANKA_094120), PTEX150 (PbANKA_100850), EXP2 (PbANKA_133430), PTEX88 (PbANKA_094130) and TRX2 (PbANKA_135800) have been identified as members of the PTEX complex. Phenotype Phenotype analyses indicate that HSP101 is located at the PVM of blood stages. Additional information In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the parasitophorous vacuole (PV). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1333 In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the endoplasmatic reticulum (ER). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1332 Other mutants See link for other HSP101 mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0931200

Gene Model P. falciparum ortholog

PF3D7_1116800

Gene product

heat shock protein 101 | chaperone protein ClpB2

Gene product: Alternative name

HSP101

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Details of the genetic modification

Name of the tag

mCherry-3xMyc

Details of tagging

C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct

(Linear) plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used

Modified PlasmoGEM construct/vector used

Plasmid/construct map

Plasmid/construct sequence

Restriction sites to linearize plasmid

Selectable marker used to select the mutant parasite

hdhfr/yfcu

Promoter of the selectable marker

pbdhfr

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

5-fluorocytosine (5-FC)

Additional remarks genetic modification

The hdfr-yfcu drug-selectable marker has been removed by negative selection with 5-FC

Additional remarks selection procedure

The hdfr-yfcu drug-selectable marker has been removed by negative selection with 5-FC

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

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