RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5643

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1344400; Gene model (P.falciparum): PF3D7_1329400; Gene product: AMP deaminase (AMPD)
Phenotype	No phenotype has been described

Last modified: 21 September 2025, 10:50

*RMgm-5643

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene disruption
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 31132185
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	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Nagappa LK, Balaram H
Name Group/Department	Molecular Biology and Genetics Unit
Name Institute	Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR)
City	Bengaluru
Country	India
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Name of the mutant parasite
RMgm number	RMgm-5643
Principal name	Δampd
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Not different from wild type
Gametocyte/Gamete	Not different from wild type
Fertilization and ookinete	Not different from wild type
Oocyst	Not different from wild type
Sporozoite	Not different from wild type
Liver stage	Not different from wild type
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of AMPD Protein (function) Phenotype Evidence is presented for the following: - AMPD is non-essential during intra-erythrocytic stages in P. berghei. Comparison of blood-stage growth rate measurements between wildtype and Δampd P. berghei parasites in mouse showed no significant difference - Enumeration of gametocytes did not show any significant difference in the number of gametocytes as well as the ratio of female to male gametocytes between wildtype and Δampd parasites. - Δampd P. berghei parasites undergo normal development in A. stephensi and produce infectious sporozoites. - Δampd P. berghei sporozoites exhibit normal gliding motility and transform into EEFs in HepG2 cells - Δampd P. berghei sporozoites initiate a timely blood stage infection in C57BL6 mice Additional information Other mutants

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1344400

Gene Model P. falciparum ortholog

PF3D7_1329400

Gene product

AMP deaminase

Gene product: Alternative name

AMPD

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Details of the genetic modification

Inducable system used

Additional remarks inducable system

Type of plasmid/construct used

(Linear) plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used

Yes

Name of PlasmoGEM construct/vector

Modified PlasmoGEM construct/vector used

Plasmid/construct map

Plasmid/construct sequence

Restriction sites to linearize plasmid

Partial or complete disruption of the gene

Complete

Additional remarks partial/complete disruption

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

The library clone for P. berghei AMPD (PbG02_B-44h05) was obtained from PlasmoGem. The procedures for knockout construct generation were described earlier (Godiska et al., 2009; Pfander et al., 2011). The intermediate vector and final knockout construct were confirmed by PCR and restriction digestion. The final knockout construct was subjected to NotI digestion, purified and used for transfection,
Δampd P. berghei parasites were generated using linear pJAZZok vector with homology arms flanking the marker gene, generated by recombineering strategy (Godiska et al., 2009; Pfander et al., 2011). The knockout parasites were genotyped by PCR and Southern blotting , and by limiting dilution three clonal populations (C1, C2 and C3) were obtained that were genotyped by PCR.

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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