RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-248
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Last modified: 15 March 2012, 10:03
*RMgm-248| 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 | 3 |
| Reference (PubMed-PMID number) | Not published (yet) |
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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 cl15cy1 |
| Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | D.L. Gardiner, C.J. Janse, A.P. Waters |
| Name Group/Department | Malaria Biology Laboratory |
| Name Institute | Queensland Institute of Medical Researchal Center |
| City | Herston |
| Country | Australia |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0836300 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0935800 | ||||||||||||||||||||||||
| Gene product | cytoadherence linked asexual protein 9 | ||||||||||||||||||||||||
| Gene product: Alternative name | CLAG9 | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
| Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| 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 | Two P. berghei gene models: PB000829.03.0, PB000048.01.0 The following target regions were used in the construct to disrupt CLAG9: Insert No1. In Hind III site 5'-3' 1 CTAAATCATAATGCCACTGTAGAAGATGTAATAAATTATGAACATATTTTAAAAGAACAA 61 GTGGCTTTAGAATACGACCCTTCGATATCAGATAAAATTAAAAATAAAATATTAATTGTG 121 AGAACTCTAAAAATTGTTAAGTTTATGTTGGTACCAATTTATGAGTTTAAAAAAAATAAA 181 GACATAAAACAAGCCTTATACAAATTAAATAATTTTTTTATTTATGATCATAAAAAAAGT 241 AAAAATCATGATTATTCCACTTCTAAATTAAAAAATGTAATTCGTGAAAATATGCTATAT 301 TCATCAAAATTTCTCAAAGTCAAAAATCAAATACCCCAAAATACCGATATCAATGAATAT 361 AATAATGACTACAAAGAAATATTATTTTCTTATGAACCTGATATTGAATATATGAAAAAC 421 CAAGATGCTATATCTAATTATTATGACATAGGCATATATAATCATATCGGAGCCCATTTA 481 TAAGTGCGCTATACATTTTCTGCATTCCCCATTTTGGCATTTCTACCATTTCTGTCATTT 541 TCGCCCCTTTTCGTTTTGCAGCGCTTGGTCATTTTATCACCCTCAAAATGGCATTCAAAC 601 ATTTCAAAAAATACTTTGTAAATGGAGATCTACGGGTTTATC and Insert No2 3' EcoR1 site. 5'-3' 1 CAAATGCTATGTTTCTAAATCTATATAATGAAACTAAACCCAAACTAACTATGATAACAT 61 TCCATTGGTACCCATCATATTTAAAGAAATTTATTATTCATTATGTTCGAAAAAATAAAG 121 CTGCATCTTTGTTAAATGGTAAATTTTAAAAATGACAAAATAATTTTTTTTTTTTTTTGC 181 ATTTCTTATATTCATCGGATTAATATTCACTCTTTTATCCATTTACAGAATTAGAAAGTA 241 AAGTAAAAAAAGAAACAATCGAAAAAATGAAAAATAGCATACGATTCGTTATGCACATTA 301 ATTTCAATATTACAATTAGACTTTTTTTATTATCTAAATGAAAAGCCTTTGAAAAATAGT 361 CATCCATATGGATTAACAATGCTAATTGAAAATAAATTTAAAGATTGGTTTATGAATTAC 421 CTAACAGGGTTTACTATGGTTAATTATGATGATAATAAAGGGCGTTATAATTTACCTGGA 481 AAACGTGAAAGGAGGGAATTTTTAGCATCAAAATTAAAAATGTGGACTTTATTTGTGAAA 541 AAAATTATAACTGATGCATATGTTCAAAACTTTAATCAAAAACATGTTATAAATTTATAT 601 AAATATCATGACATATTCAATATAAATAACAAAATTATGGTAATGAGGGATTCCTATGAA 661 CTTTATATC Disruption experiments were performed in the Leiden Malaria Research Group (exp. 305, 306, 319) RMgm-723 describes an independent attempt to disrupt PBANKA_083630 | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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