RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-16

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_0524800; Gene model (P.falciparum): Not available; Gene product: early transcribed membrane protein small exported protein (SEP1; ETRAMP; Pbsep1)
Phenotype	No phenotype has been described

Last modified: 28 August 2011, 22:26

*RMgm-16

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) : 10984459 Reference 2 (PMID number) : 12615320
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	8417HP
Other information parent line	A reference wild type clone from the ANKA strain of P. berghei (Janse et al., Exp. Parasitol. 68, 274-282).
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The mutant parasite was generated by
Name PI/Researcher	Pace, T; Ponzi, M
Name Group/Department	Laboratorio di Biologia Cellulare
Name Institute	Istituto Superiore di Sanitá
City	Rome
Country	Italy
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Name of the mutant parasite
RMgm number	RMgm-16
Principal name	HP∆5
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 SEP1 (early transcribed membrane protein, ETRAMP). Protein (function) In P. berghei, this gene belongs to a family of three related genes (encoding so-called SEP proteins; SEP1-3), located in the subtelomeric regions of chromosomes 5 (Birago et al., Mol. Biochem. Parasitol. (2003) 126, 209-18). The SEP proteins show homology to members of the ETRAMP family of proteins of P. falciparum but the orthologous relationship of the different members is unclear (ortholog of etramp11.1, PF11_0039?). The sep1 gene encodes encodes an integral membrane protein that is expressed during the entire erythrocytic cycle and is present in the parasitophorous vacuole membrane. The protein contains a predicted signal peptide at the NH(2)-terminus, an internal hydrophobic region and a polymorphic, low-complexity region at the carboxy-terminus (Birago et al., Mol. Biochem. Parasitol. (2003) 126, 209-18). Phenotype The lack of expression of SEP1 does not result in a clear phenotype. The phenotype of the mosquito and liver stages has not been analysed in detail but the mutant parasites can esaily be transmitted by Anopheles stephensi (pers. comm. Dr. M. Ponzi). See also the mutant parasite line RMgm-18. In this mutant the same gene has been dirupted without a clear phenotype. Additional information Sep1 in the mutant described here (named orfA or SEP1) has been deleted from the genome by a method that induces terminal deletions at specific chromosome ends, using a linear construct containing telomeric sequences at one end and target sequences of the gene at the other to drive homologous recombination (Pace et al., Genome Res. (2000) 10, 1414-20). Other mutants An independent mutant lacking expression of SEP1 has been generated (see RMgm-18) The two other sep genes, sep2 (PB106367.00.0; early transcribed membrane protein, ETRAMP; sep-bis) and sep3 (PB401867.00.0; sep-ter) have been unsuccessfully targeted for gene disruption (pers. comm. M. Ponzi, T. Pace). See RMgm-64 (sep2) and RMgm-65 (sep3).

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0524800

Gene Model P. falciparum ortholog

Not available

Gene product

early transcribed membrane protein small exported protein

Gene product: Alternative name

SEP1; ETRAMP; Pbsep1

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Details of the genetic modification

Inducable system used

Additional remarks inducable system

Type of plasmid/construct used

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

Partial or complete disruption of the gene

Complete

Additional remarks partial/complete disruption

Selectable marker used to select the mutant parasite

pbdhfr

Promoter of the selectable marker

pbdhfr

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

This gene (named orfA) was deleted from the genome by a method that induces terminal deletions at specific chromosome ends, using a linear construct containing telomeric sequences at one end and target sequences of the gene at the other to drive homologous recombination (Pace et al., Genome Res. (2000) 10, 1414-20).

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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