RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-5089
|
||||||||||
Last modified: 26 October 2021, 17:11
*RMgm-5089| Successful modification | The gene/parasite could not be changed/generated by the genetic modification. |
| The following genetic modifications were attempted | Gene disruption |
| Number of attempts to introduce the genetic modification | Unknown |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 34604117 |
| top of page | |
| Parent parasite used to introduce the genetic modification | |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone | Not applicable |
| Other information parent line | |
| top of page | |
| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Niikura M, Kobayashi F |
| Name Group/Department | Department of Environmental Science, School of Life and Environmental Science |
| Name Institute | Azabu University |
| City | Kanagawa |
| Country | Japan |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1232100 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0517300 | ||||||||||||||||||||||||
| Gene product | serine/arginine-rich splicing factor 1 | ||||||||||||||||||||||||
| Gene product: Alternative name | sr1 | ||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||
| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | (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 | |||||||||||||||||||||||||
| Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| 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 | The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood-stage growth/multiplication. Nuclear poly(A) binding protein 2 (NAB2), THO complex subunit 4 (THO4), nucleolar protein 3 (NPL3), G-strand binding protein 2 (GBP2) and serine/arginine-rich splicing factor 1 (SR1) are involved in nuclear mRNA export in malaria parasites. In S. cerevisiae, individual serine/arginine-rich (SR) proteins, such as NPL3, GBP2, and HRB1 are not essential. NPL3, GBP2 and SR1 contain conserved RNA recognition motifs. To generated nab2-, tho4-, npl3- and sr1-deleted P. berghei ANKA, the gene-targeting vectors for nab2 (PBANKA_1122000), tho4 (PBANKA_1230500), npl3 (PBANKA_0506600) and sr1 (PBANKA_1232100) were prepared by PCR. Briefly, the 5’ and 3’ flanking regions of the open reading frame (ORF) of target genes were amplified by PCR. The PCR products were annealed to either side of the human dihydrofolate reductase (hdhfr)-expressing cassette and amplified by PCR using gene-specific primers The gene-targeting vectors were introduced into the ORFs of target genes by double-crossover homologous recombination | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
|
Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
| |||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||