RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-237

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1024600; Gene model (P.falciparum): PF3D7_1418100; Gene product: liver specific protein 1, putative (LISP1, liver specific protein 1)
Phenotype	Liver stage;

Last modified: 23 May 2009, 23:21

*RMgm-237

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) : 19438514
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei NK65
Name parent line/clone	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	T. Ishino, P. Baldacci
Name Group/Department	Biologie et Génétique du Paludisme
Name Institute	Institut Pasteur
City	Paris
Country	France
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Name of the mutant parasite
RMgm number	RMgm-237
Principal name	NK65Lisp1Δ
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	By analysis of blood stage parasitemia after intravenous injection of sporozoites, the mutant sporozoites showed a 15-fold decrease in infectivity.
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of LISP1 (liver specific protein 1). Protein (function) Phenotype analyses of liver stage development of this and additional mutants lacking expression of LISP1 (RMgm-279, RMgm-280) indicate that LISP plays a role in egress of merozoites from liver cells. Phenotype Phenotype analyses of liver stage development of additional mutants lacking expression of LISP1 (RMgm-279, RMgm-280) indicate that LISP plays a role in egress of merozoites from liver cells, resulting from a defect in rupture of the membrane of the parasitophorous vacuole (PVM) and remain trapped inside hepatocytes. Additional information Using polyclonal antibody expression was detected in 48h liver schizonts in livers from rats and in infected HepG2 cells at 36, 48 and 65h post infection. No signal was detected in blood stages and sprorozoites. IFA analysis indicated a location of LISP in the parasitophorous vacuole membrane (PVM). Four gene models exist: PB000708.00.0; PB000682.00.0; PB001247.00.0; PB000250.00.0. Gen Bank accession no: AB231328. Other mutants RMgm-279: An independent mutant lacking expression of LISP1 and expressing GFP under the constitutive eef1a promoter RMgm-280: An independent mutant lacking expression of LISP1 (ANKA strain)

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1024600

Gene Model P. falciparum ortholog

PF3D7_1418100

Gene product

liver specific protein 1, putative

Gene product: Alternative name

LISP1, liver specific protein 1

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Details of the genetic modification

Inducable system used

Additional remarks inducable system

Type of plasmid/construct used

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

Partial or complete disruption of the gene

Complete

Additional remarks partial/complete disruption

Selectable marker used to select the mutant parasite

hdhfr

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	5’- CGATGCGGGCCCGAGAATACAACTTACTAGGAAATGCAC-3’
Additional information primer 1	ApaI - 630bp
Sequence Primer 2	5’- AGCTGGCGCGCCCTTGCATCTTCAGACATGTTATTTTCGA-3’
Additional information primer 2	SmaI - 630bp
Sequence Primer 3	5’- CCAGTGAGTGCGGCCGCGGCTAAAGCTCGATGTCGTATTCAAGAA-3’
Additional information primer 3	NotI - 600bp
Sequence Primer 4	5’- AGCTGGCGCGCCCTGGGAATTTTTCAATTTCTTCCATTTG-3’
Additional information primer 4	AscI - 600bp
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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