RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4672

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_0716000; Gene model (P.falciparum): PF3D7_0414000; Gene product: structural maintenance of chromosomes protein 3, putative (SMC3) Name tag: triple-HA
Phenotype	Asexual bloodstage;

Last modified: 18 August 2019, 14:06

*RMgm-4672

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) : 31390575
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	Hillier C, Billker O, Choudhary JS
Name Group/Department	Functional Proteomics
Name Institute	The Institute of Cancer Research
City	London
Country	UK
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Name of the mutant parasite
RMgm number	RMgm-4672
Principal name	PBANKA_0716000-3xHA
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Expression in schizonts. Epitope-tagged PBANKA_0716000 was immunoprecipitated from the lysates of P. berghei schizonts, and MS identification of co-purifying proteins confirmed interaction with different proteins.
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 C-terminal 3xHA-tagged version of PBANKA_0716000 Protein (function) structural maintenance of chromosomes protein 3, putative Phenotype The protein was identified in a large-scale protein interaction network analysis. A physical association was confirmed between putative cohesin subunits SMC3 (PBANKA_0716000) and SMC1 (PBANKA_0917500) and identified a further SMC3-associated protein, uncharacterized protein PBANKA_1304000, which in the network is connected to both SMC1 and SMC3 through another uncharacterized protein that was not detected in the pull-downs. Additional information Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0716000

Gene Model P. falciparum ortholog

PF3D7_0414000

Gene product

structural maintenance of chromosomes protein 3, putative

Gene product: Alternative name

SMC3

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Details of the genetic modification

Name of the tag

triple-HA

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

Yes

Name of PlasmoGEM construct/vector

PbGEM-281970

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

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

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

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