SummaryRMgm-5196
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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) : 35404079 |
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 | Wang PP, Zhu Z |
Name Group/Department | Department of Immunology, College of Basic Medical Sciences |
Name Institute | China Medical University |
City | Shenyang |
Country | China |
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Name of the mutant parasite | |
RMgm number | RMgm-5196 |
Principal name | Δpbgpr180(del) |
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 | Normal (wild type) numbers of gametocytes are produced; male gamete formation ~2fold reduced (as determined by counting exflagellation); female gamete formation reduced by ~ 30%. Crossing experiments indicate a more severe defect in female gametes compared to males. |
Fertilization and ookinete | Reduced ookinete formation (Whereas >93% of wild-type zygotes were transformed into mature ookinetes after 24 h of culture, Δpbgpr180 parasites had a ~50% conversion rate. |
Oocyst | ~50% reduction in the percentage of infected mosquitoes and ~80% decrease in oocyst density. |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Phenotype To investigate the function of GPR180i, we generated two pbgpr180 knockout (KO) lines Δpbgpr180(Del) (with the PbGEM-340051 plasmid) and Δpbgpr180KO (with the pL0034-PbGPR180-KO plasmid) using a double-crossover homologous recombination strategy. Crossing experiments indicate a more severe defect in female gametes compared to males. Evidence is presented that: Other mutants |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1429300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1213500 | ||||||||||||||||||||||||
Gene product | integral membrane protein GPR180, putative | ||||||||||||||||||||||||
Gene product: Alternative name | GPR180 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) PCR construct double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | Yes | ||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | PbGEM-340051 | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | For pbgpr180 gene disruption, two plasmids were used. The PbGEM-340051 plasmid (to delete nt: 392 – 2080 bp of pbgpr180 open reading frame–ORF) was obtained from PlasmoGEM. To delete the entire pbgpr180 ORF, an 1157 bp fragment containing the 5’UTR (nt: 21157 – 21 bp) and the 871 bp 3R described above were amplified and cloned into the HindIII/ApaI and XhoI/NotI sites, respectively, of the pL0034 vector to generate the plasmid pL0034-PbGPR180-KO. For transfection, the pL0034-PbGPR180-KO plasmid was linearized with BglI and NotI, while the PbGEM-340051 plasmid was linearized with NotI, followed by ethanol precipitation | ||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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