Back to search resultsSummaryRMgm-148
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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) : 15533999 |
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 | E.I. khater; R.E. Sinden; J.T. Dessens |
Name Group/Department | Department of Biological Sciences |
Name Institute | Imperial College |
City | London |
Country | United Kingdom |
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Name of the mutant parasite | |
RMgm number | RMgm-148 |
Principal name | IMC1a-KO (clone 104, 204) |
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 | Oocysts appeared to develop normally and formed large numbers of sporozoites. However, closer examination of these sporozoites revealed that they were of abnormal shape and some 20–30% smaller in size. Each sporozoite possessed a single enlarged, protruding area associated with the position of the nucleus. The position of these protrusions varied between sporozoites from being located at the posterior end to being positioned near the middle of the sporozoite. The mutant sporozoites accumulated in the hemolymph and did not invade the salivary glands. Gliding motility was strongly reduced. Mutant sporozoites were able to attach normally to the glass surface and undergo circular gliding, but their gliding speed was fivefold slower than that of wild type sporozoites. Inoculation of purified oocyst-derived/hemolymph sporozoites into mice did not result in infection of the mice. |
Liver stage | Inoculation of purified oocyst-derived/hemolymph sporozoites into mice did not result in infection of the mice. |
Additional remarks phenotype | Mutant/mutation Protein (function) Additional information |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0402600 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0304000 | ||||||||||||||||||||||||
Gene product | inner membrane complex protein 1a, putative | ||||||||||||||||||||||||
Gene product: Alternative name | IMC1a, inner membrane complex protein 1a | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | 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 | KpnI/NotI | ||||||||||||||||||||||||
Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | A modified T. gondii dihydrofolate reductase/thymidylate synthase (DHFR/TS) gene cassette (conferring resistance to pyrimethamine) was inserted into the imc1a gene by double homologous recombination replacing nucleotides 550–2,404 of the imc1a gene | ||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
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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