RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1423

Malaria parasite	P. chabaudi
Genotype
Tagged	Gene model (rodent): PCHAS_1100200; Gene model (P.falciparum): Not available; Gene product: CIR protein; PIR protein Name tag: GFP
Phenotype	Asexual bloodstage;

Last modified: 30 March 2016, 13:53

*RMgm-1423

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) : 26996203
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. chabaudi
Parent strain/line	P. c.chabaudi AS
Name parent line/clone	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Yam XY; Langhorne J; Preiser PR
Name Group/Department	School of Biological Sciences
Name Institute	Nanyang Technological University
City	Singapore
Country	Singapore
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Name of the mutant parasite
RMgm number	RMgm-1423
Principal name	PCHAS_110020::GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	GFP expression was observed in the parasitophorous vacuole and in the RBC cytoplasm in trophozoites and associated with merozoites in schizonts.
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 GFP-tagged version of the protein. The transgene is introduced on a plasmid and is expressed under the control of the constitutive eef1a promoter of P. berghei Protein (function) The protein is a member of a variant antigen family (Plasmodium interspersed repeat proteins, PIR). These proteins are exported into host erythrocyte. In P. vivax there are approximately 350 pir (vir) in the genome, and homologues have been identified in Plasmodium infecting rodents and monkeys; P. berghei (bir), P. chabaudi (cir), P. yoelii (yir), P. knowlesi (kir). PIR proteins show homology to proteins which are members of the RIF/STEVOR family of P. falciparum proteins. Phenotype This study analysed the localisation of 11 members of the P. c. chabaudi (AS) PIR (CIR) multigene family in mutants that expressed a GFP-tagged version from an episome and under control of the constitutive eef1a promoter from P. berghei. GFP expression was observed in the parasitophorous vacuole and in the RBC cytoplasm in trophozoites and associated with merozoites in schizonts. See the link CIR::GFP for the other 10 mutants 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

PCHAS_1100200

Gene Model P. falciparum ortholog

Not available

Gene product

CIR protein; PIR protein

Gene product: Alternative name

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Details of the genetic modification

Name of the tag

GFP

Details of tagging

C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct

Circular plasmid

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

tgdhfr

Promoter of the selectable marker

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

Full length coding sequences of cir genes were generated by chemical gene synthesis (GeneScript) and were amplified by PCR for cloning into plasmid ePL vector with GFP tag under a constitutive promoter (P. berghei ef1a) (in house made vector.

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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