RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4315

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1328900; Gene model (P.falciparum): PF3D7_1465500; Gene product: potassium channel K2 (KCh2, K2)
Phenotype	No phenotype has been described

Last modified: 7 September 2017, 14:13

*RMgm-4315

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) : 28864420
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	P. berghei ANKA 2.34
Other information parent line	P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
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The mutant parasite was generated by
Name PI/Researcher	P. Ellekvist; N. Kumar; DA Klaerke
Name Group/Department	Department of Physiology and Biochemistry, IBHV, Faculty of Life Sciences
Name Institute	University of Copenhagen
City	Copenhagen
Country	Denmark
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Name of the mutant parasite
RMgm number	RMgm-4315
Principal name	kch2-null
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 tested
Fertilization and ookinete	Not tested
Oocyst	Not different from wild type
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of KCh2 (potassium channel K2) Protein (function) K+ channels constitute the largest and most diverse of ion channel families and are involved in K+ transport, cell volume control, and regulation of membrane potential. Two putative K+ channel-encoding genes have been found in the Plasmodium falciparum genome (Kch2; PBANKA_1442000; PF3D7_1227200) Phenotype Normal blood stage growth/multiplication and formation of oocysts Additional information Evidence is presented for: '⁸⁶Rb⁺ uptake in Kch2-deficient blood-stage P. berghei parasites (Kch2-null) did not differ from that of wild-type (WT) parasites. About two-third of the ⁸⁶Rb⁺ uptake in WT and in Kch2-null parasites could be inhibited by K+ channel blockers and could be inferred to the presence of functional Kch1 in Kch2 knockout parasites. Kch2 is therefore not required for transport of K+ in P. berghei and is not essential to mosquito-stage sporogonic development of the parasite'. Other mutants PF3D7_1227200 : KCh1 mutants PF3D7_1465500 : KCh2 mutants

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1328900

Gene Model P. falciparum ortholog

PF3D7_1465500

Gene product

potassium channel K2

Gene product: Alternative name

KCh2, K2

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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

Modified PlasmoGEM construct/vector used

Plasmid/construct map

Plasmid/construct sequence

Restriction sites to linearize plasmid

KpnI, NotI, and ScaI

Partial or complete disruption of the gene

Partial

Additional remarks partial/complete disruption

Targeting plasmid (pB3DPbKch2) was constructed by cloning fragments from the 5’ and 3’ flanking regions of the deduced six transmembrane domains-encoding part of the PbKch2 gene

Selectable marker used to select the mutant parasite

tgdhfr

Promoter of the selectable marker

pbdhfr

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
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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