SummaryRMgm-4924
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene tagging |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 33222390 |
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 | Liu F, Cao Y |
Name Group/Department | Department of Immunology, College of Basic Medical Sciences |
Name Institute | China Medical University |
City | Shenyang, Liaoning |
Country | China |
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Name of the mutant parasite | |
RMgm number | RMgm-4924 |
Principal name | Pb22-HA |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Immunofluorescence analyses showed the following: In the erythrocytic stages, Pb22 expression was restricted to the parasites in schizonts and gametocytes. Abundant expression of the Pb22 protein in both male and female gametocytes. |
Gametocyte/Gamete | Immunofluorescence analyses showed the following: In the erythrocytic stages, Pb22 expression was restricted to the parasites in schizonts and gametocytes. Abundant expression of the Pb22 protein in both male and female gametocytes. Evidence presented for surface location in gametes and ookinetes. |
Fertilization and ookinete | Immunofluorescence analyses showed the following: In the exflagellating male gametes, the protein was observed on the residual body as well as the flagella. Evidence presented for surface location in gametes and ookinetes. |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Phenotype Immunofluorescence Analyses of the mutant expressing the HA-tagged Pb22 showed the following: In the erythrocytic stages, Pb22 expression was restricted to the parasites in schizonts and gametocytes. Abundant expression of the Pb22 protein in both male and female gametocytes. In the exflagellating male gametes, the protein was observed on the residual body as well as the flagella. Evidence presented for surface location in gametes and ookinetes. Additional information Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0305900 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0208800 | ||||||||||||||||||||||||||
Gene product | conserved Plasmodium protein, unknown function | ||||||||||||||||||||||||||
Gene product: Alternative name | pb22 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | HA | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid double cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | To tag the endogenous Pb22 with an HA tag, the 5′ fragment was amplified with the primers HA-5UTR-F and HA-5UTR-R, while the 3′ fragment was amplified with primers HA-3UTR-F and HA-3UTR-R. The fragments were cloned into vector PL0035 (containing the human dhfr cassette, yfcu cassette and ha tag) | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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