RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4126

Malaria parasite	P. berghei
Genotype
Genetic modification not successful
Mutated	Gene model (rodent): PBANKA_0926700; Gene model (P.falciparum): PF3D7_1121600; Gene product: exported protein 1 \| circumsporozoite-related antigen \| parasitophorous vacuole membrane antigen QF 116 (EXP1; CRA; Ag5.1; HEP17) Details mutation: Replacement of the promoter region by the ama1-PBANKA_091500-promoter by 'promoter swap' technology
Phenotype	No phenotype has been described

Last modified: 5 February 2017, 11:09

*RMgm-4126

Successful modification	The gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted	Gene mutation
Number of attempts to introduce the genetic modification	Unknown
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 28115054
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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	Not applicable
Other information parent line
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Attempts to generate the mutant parasite were performed by
Name PI/Researcher	Klug D, Frischknecht F
Name Group/Department	Integrative Parasitology
Name Institute	Center for Infectious Diseases, Heidelberg University Medical School
City	Heidelberg
Country	Germany

Mutated: Mutant parasite with a mutated gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0926700

Gene Model P. falciparum ortholog

PF3D7_1121600

Gene product

exported protein 1 | circumsporozoite-related antigen | parasitophorous vacuole membrane antigen QF 116

Gene product: Alternative name

EXP1; CRA; Ag5.1; HEP17

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Details of the genetic modification

Short description of the mutation

Replacement of the promoter region by the ama1-PBANKA_091500-promoter by 'promoter swap' technology

Inducable system used

Short description of the conditional mutagenesis

Not available

Additional remarks inducable system

Type of plasmid/construct

(Linear) plasmid single 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

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

Unsuccessful attempts to disrupt the exp1 indicates an essential function (see RMgm-4125).
Also replacement of the exp1 promoter region with the (schizont-specific) ama1 promoter region were unsuccessful.

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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