Summary

RMgm-1448
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1003000; Gene model (P.falciparum): PF3D7_0405300; Gene product: liver specific protein 2, putative | sequestrin | 6-cysteine protein (LISP2)
Phenotype Liver stage;
Last modified: 2 June 2016, 18:18
  *RMgm-1448
Successful modificationThe 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) : 27241521
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherKumar H; Frischknecht F
Name Group/DepartmentIntegrative Parasitology
Name InstituteCenter for Infectious Diseases, Heidelberg University Medical School
CityHeidelberg
CountryGermany
Name of the mutant parasite
RMgm numberRMgm-1448
Principal nameLISP2(–)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNormal numbers of sporozoites are formed that are infective to cultured hepatocytes. Parasites arrest during development in hepatocytes after a period of growth. In most parasites no merozoites are formed as shown by the low level of MSP1 expression. After intravenous injection of sporozoites, 1 out of 60 mice developed a blood stage infection
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of LISP2

Protein (function)
PBANKA_100300 encodes a 2172 amino acid protein with a large repeat region of approximately 1000 amino acids. This region is composed of several repeat motifs, particularly 12 copies of a 56-amino acid motif. Analyses of the amino acid sequence with various structure analysis tools indicates that it has an N-terminal signal sequence but no other motifs for membrane association such as transmembrane domains or a GPI-anchor motif, suggesting that the encoded product has the structure of a secreted protein. It contains a predicted Plasmodium 6-cysteine motif. Several studies analysing expression of this protein in P. berghei (see 'Additional mutants') provide evidence for specific expression in liver stages. The protein has been named LISP2 and sequestrin.

Phenotype
Normal numbers of sporozoites are formed that are infective to cultured hepatocytes. Parasites arrest during development in hepatocytes after a period of growth. In most parasites no merozoites are formed as shown by the low level of MSP1 expression. After intravenous injection of sporozoites, 1 out of 60 mice developed a blood stage infection (see also below).

Additional information
The mutant described here show a stronger growth-arrested phenotype in comparison to earlier described mutants lacking expression of LISP2 (see RMgm-799, RMgm-930, RMgm-931), possibly due to expression of the N-terminal region of the lisp2 gene in the latter mutants. Evidence is presented that expression of a small part of the N-terminal region of lisp2 in parasites lacking lisp2 results in a reduced growth arrest in the liver.

Other mutants
See the link PBANKA_1003000 for other related mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1003000
Gene Model P. falciparum ortholog PF3D7_0405300
Gene productliver specific protein 2, putative | sequestrin | 6-cysteine protein
Gene product: Alternative nameLISP2
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
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
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