Summary

RMgm-221
Malaria parasiteP. berghei
Genotype
Transgene
Transgene Plasmodium: Gene model: PBANKA_0837161; Gene model (P.falciparum): Not available; Gene product: BIR protein PIR protein (BIR38, Plasmodium berghei interspersed repeat protein 38)
Promoter: Gene model: PBANKA_0837161; Gene model (P.falciparum): Not available; Gene product: BIR protein PIR protein (BIR38, Plasmodium berghei interspersed repeat protein 38)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Insertion locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c-type unit))
Phenotype Asexual bloodstage;
Last modified: 8 April 2009, 10:40
  *RMgm-221
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 18563749
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
The mutant parasite was generated by
Name PI/ResearcherF. Di Girolamo, C.J.Janse, M. Ponzi
Name Group/DepartmentDipartimento di Malattie Infettive Parassitarie ed Immunomediate
Name InstituteIstituto Superiore di Sanità
CityRome
CountryItaly
Name of the mutant parasite
RMgm numberRMgm-221
Principal name580cl1
Alternative nameBir38-gfp (ssu-rrna)
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageBIR38-GFP expression in asexual blood stages. BIR38-GFP is found in cytoplasm of infected erythrocytes and is associated with cholesterol-rich microdomains (lipid rafts). During the asexual blood stage cycle of 22-24 hour, GFP expression was detected in late trophozoites and developing schizonts.
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a GFP-tagged (c-terminal) form of BIR38. The bir38-gfp gene is introduced into the non-essential c-type ribosomal rna gene and is expressed under the control of its own 5'-UTR and pbdhfr 3'-UTR regulatory sequences.

Protein (function)
BIR38 is a member of a variant antigen family (Plasmodium interspersed repeat proteins, PIR). These proteins are potentially implicated in host immune system interactions and targeted to or on the surface of the host erythrocyte. In P. vivax there are approximately 350 pir (vir) in the genome, and homologues have been identified in Plasmodium infecting rodents and monkeys; P. berghei (bir), P. chabaudi (cir), P. yoelii (yir), P. knowlesi (kir). PIR proteins show homology to proteins which are members of the RIF/STEVOR family of P. falciparum proteins. Consistent with a role in antigenic variation and immune evasion, PIR proteins have been demonstrated on, or close to the surface membrane of erythrocytes infected with P. vivax, P. chabaudi, P. yoelii and P.berghei  and switching of yir transcription has been observed in response to a host immune response.

Phenotype
The phenotype analyses indicate that BIR38 is expressed late during the asexual blood stage cycle (late trophozoites, schizonts) and is exported to the host erythrocyte cytoplasm and is associated with cholesterol-rich microdomains (lipid rafts).

Additional information
In plasma membranes of eukaryotic cells associations of sphingolipids, cholesterol and proteins enable the formation of special microdomains that are resistant to solubilization with non-ionic detergents at low temperature. These detergent-resistant membranes, named lipid rafts, play a role in defining the specificity of trafficking of proteins and lipids to and from cellular compartments. Many raft proteins are linked to saturated acyl chains in the form of glycosylphospatidylinositol (GPI) anchor or through acylation with myristate or palmitate.

DNA sequence (unspliced) of bir38 (introns in blue):
ATGGATGATG CTCTA GTATA TACACTTTTT TATATTATAT TTCCTATAAA CTTCATTAAG TTTTATTTTA ATATCATTTT CTTAATACAT ATTTTTACCA TTTTTTTTAA ATGGTTTCTT AGTGTCAAAA CTTTGATGTT TTGATGAAGT ATTTACCTGC TGAATTAGGC GGAACCGCAA ATTTTGAATT TAAACAAATT ACAAATTTCG ATAAGTACTG CCCTAATGGA GACTGCAATA CTAATCTCGA AAAAATTACA ATTGGATTTT TATGGTTACT TGAACAATGT TTTTCTACAC CCATAAATAA AAATCATAAT GAAAACAATA CTAATGCATA TTTTATATAT GTTGTATTAT GGTTAAGTTA CAAATTAAAT CAAATAACAG GCCAAAATTT CACTAAAATA AACGATTTTT ATACTAATCA AGTAAATAAT AGTGGTAAAT ATAGTAAATT TATAAGTGAT TCCAATAGAT ATACAAATAT TAAGGAATTC ATAGAGAAAC AAAAAGATTT GTTGAATATT AATATTGAAG ATCTTTCTAA ATTTTATGAT GCATCCAAAT TAATATGTAG TATGTATGGT AATTATGCAA TGAATAATAA TAACAAACTG TCAAATAATG CTACTAGTTT TGTTAACACA TATACAGAGC TTAACAATAA CTATAATACT AAAGGTACCG CATATAGTCA AATACTGTCT GCCTTATCAA CTGATTATAA TAATTTAAAA AATAAATGTA GCAATATTCC AACCCTTTCA GAGATAAACA CATCAAAAAA TACTGTAGAA ACTTCTGATC AAAGTCCTGA ACAAAGTGTA CAAATATTAG AACATATTTC TGAAGTTACA TCATCAAGTT CGTCGATAGG AAACAAATTA TTTACAGTTT TATCGATATT TGGTGCAATA GCATTTTTTT TAGGAATTTC TTATAAGGTA AATAATAAGG AATTAAAAAA ATATTTTCAT TATATATATG CAAGCATTAA CAAAAATACC ATACGCTTCT TAACGTTTTA TATTAGTATT CGTTATTTGG ACGTCGGAAA CGATCTCAAA AACAATATTT AAGAGAAAAA ATAAAAAATA TAAAGAAGAG AATGAATCAT TAA

Protein BIR38 (transmembrane region in pink):
MDDALCQNFD VLMKYLPAEL GGTANFEFKQ ITNFDKYCPN GDCNTNLEKI TIGFLWLLEQ CFSTPINKNH NENNTNAYFI YVVLWLSYKL NQITGQNFTK INDFYTNQVN NSGKYSKFIS DSNRYTNIKE FIEKQKDLLN INIEDLSKFY DASKLICSMY GNYAMNNNNK LSNNATSFVN TYTELNNNYN TKGTAYSQIL SALSTDYNNL KNKCSNIPTL SEINTSKNTV ETSDQSPEQS VQILEHISEV TSSSSSIGNK LFTVLSIFGA IAFFLGISYK YSLFGRRKRS QKQYLREKIK NIKKRMNH

Other mutants


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: Plasmodium
Gene Model of Parasite PBANKA_0837161
Gene Model P. falciparum ortholog Not available
Gene productBIR protein PIR protein
Gene product: Alternative nameBIR38, Plasmodium berghei interspersed repeat protein 38
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid single cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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CTGAAACGCGGGAATATTATACATTGATTTAAAGTATTTTCATTAAGCGCACTAATTATT
CCGACGTACTATACAATGATATAGCACTGACAATAGATAAACGTATTAAATACGAACAAA
TCATTAAATTAATTTGATTCGGATACATTGATATATATTATTAAGCTTGTAATAAGATTA
AAATTATATGCGCAAAACATCAAATGAAATATTATAACACTTATTTAAGTAAGATTTAAT
GCGATGACATTAGACTAACACTATCAAGCAAAATAATATTAATAATTATAGTCTTCTTAA
AAATAATTTTTTTCATCATAAAACTAAATATAGTTTATTATAAAATATAAGAGCAAATTA
TATATTTAGAAAATAATTCTAAGCCATTTTTAATTTATACATTAATTGAAAGTTTGAATG
GTAAAAAATATAATTAAACTATGTAGAATATGTAAATCTTATATGGATACATCATTGTCT
TAAAAAAGCATAATTCCCTTATCTCTCCTCTCAAAGTGCAATATACCAACCTAATTACGC
AAAGTTTTCTATTATACTTAAAAATTTACATCAATACTTATTTGTTTATTAATATTTAAT
ATTTACTAAGTATTATTTTAAAAACAATATACATTAAAAGACTTATTTAAGCTTATAAAT
AACATAGTAAATACAGTTTCAGTGCTAATTCTCGTTTTAAACCGTGGAAAAGATGTGCAA
AATATATTAAAAATTATCCAATAAGGAATATTTCTAAATTAAAGTATATAGGAATACAAA
TAATGTTATAAGACTCATTAAAATGAATTATTAATTTATCTACATTAAATAAAAATAATA
TTATAATTATGTAATTTGCATAGAAAATGGAACATTCAACTTAATTTGTAAATGTTATAA
TTTTAATAAAATATTGGTATATATTATAAGAAACAAGTATATAAATATTTAATATATTTT
AAAAAAATTATATTTATATACTTTTAAAGATTATAGTAATAATAACTTTTTTAATTTTTT
ATATTTTTGCAGTATTTAAGTTTTAGGATATTGTCTATGTTCTATATTAAATCATATGTA
TAAGAAAATATATTATAAATTTATTATAAAAGTATATTAATTTTTATAATAATGCTCATA
TGAGTGTTATTAAGTATAAAAATTTATTCAAAATTGTATTATATATACATATGATAAAAC
ATGTATTAAACACCCTAAAACAAGGTAATTTATCTAACTATCATAACAAAAATATAATAT
TCTTTTAGTAATAATATTATATTATTATTACATATTAATTTTAAATTGAAATTAATAATA
ATAAAAATATTTTTATCTACAGATAATTGCTATATATAGGGTTAAATAATTGTATTACCT
ATTTTAGTATATATATAATATGATATCCCCTTAACCTAGAAATCATAAATTATATTACAT
CACAATGGATGATGCTCTAGTATATACACTTTTTTATATTATATTTCCTATAAACTTCAT
TAAGTTTTATTTTAATATCATTTTCTTAATACATATTTTTACCATTTTTTTTAAATGGTT
TCTTAGTGTCAAAACTTTGATGTTTTGATGAAGTATTTACCTGCTGAATTAGGCGGAACC
GCAAATTTTGAATTTAAACAAATTACAAATTTCGATAAGTACTGCCCTAATGGAGACTGC
AATACTAATCTCGAAAAAATTACAATTGGATTTTTATGGTTACTTGAACAATGTTTTTCT
ACACCCATAAATAAAAATCATAATGAAAACAATACTAATGCATATTTTATATATGTTGTA
TTATGGTTAAGTTACAAATTAAATCAAATAACAGGCCAAAATTTCACTAAAATAAACGAT
TTTTATACTAATCAAGTAAATAATAGTGGTAAATATAGTAAATTTATAAGTGATTCCAAT
AGATATACAAATATTAAGGAATTCATAGAGAAACAAAAAGATTTGTTGAATATTAATATT
GAAGATCTTTCTAAATTTTATGATGCATCCAAATTAATATGTAGTATGTATGGTAATTAT
GCAATGAATAATAATAACAAACTGTCAAATAATGCTACTAGTTTTGTTAACACATATACA
GAGCTTAACAATAACTATAATACTAAAGGTACCGCATATAGTCAAATACTGTCTGCCTTA
TCAACTGATTATAATAATTTAAAAAATAAATGTAGCAATATTCCAACCCTTTCAGAGATA
AACACATCAAAAAATACTGTAGAAACTTCTGATCAAAGTCCTGAACAAAGTGTACAAATA
TTAGAACATATTTCTGAAGTTACATCATCAAGTTCGTCGATAGGAAACAAATTATTTACA
GTTTTATCGATATTTGGTGCAATAGCATTTTTTTTAGGAATTTCTTATAAGGTAAATAAT
AAGGAATTAAAAAAATATTTTCATTATATATATGCAAGCATTAACAAAAATACCATACGC
TTCTTAACGTTTTATATTAGTATTCGTTATTTGGACGTCGGAAACGATCTCAAAAACAAT
ATTTAAGAGAAAAAATAAAAAATATAAAGAAGAGAATGAATCATCCCATGGTGAGCAAGG
GCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACG
GCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCC
TGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCC
TGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCT
TCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACG
GCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCG
AGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACA
ACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGA
ACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGC
AGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCC
AGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCG
TGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAGCGGCCGCGACT
CTAGAAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGT
AAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTA
AAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCT
AGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAA
TTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATT
ATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGA
TTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAA
AAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAGCTCGAATTC
AACCTGGTTGATCTTGCCAGTAGTCATATGCTTGTCTCAAAGATTAAGCCATGCAAGTGA
AAGTATATGCACATTTATTGCAGAAACTGCGAACGGCTCATTAAAACAGTTATAATCTAC
TTGACATTTTATTATAAGGATAACTACGGAAAAGCTGTAGCTAATACTTGTTAAGTACTT
TTACTCCCCGGAGTAATTGTATGTATTTGTTAAGACCCCTAAGAAAAAATGATATTAAAG
GAATTATAACAAAGAAGCAACACATAATATAATTATTCAGTGTGTATCAATCGAGTTTCT
GACCTATCAGCTTTTGATGTTAGGGTATTGACCTAACATGGCTTTGACGGGTAACGGGGA
ATTAGAGTTCGATTCCGGAGAGGGAGCCTGAGAAATAGCTACCACATCTAAGGAAGGCAG
CAGGCGCGTAAATTACCCAATTCTAAATAAGAGAGGTAGTGACAAGAAATAACAATATAA
GGCCAAATTTTGGTTTTATAATTGGAATGATGGGAATTTAAAACCTTCCCAAAAATCAAT
TGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAA
ATTGTTGCAGTTAAAACGCTCGTAGTTGAACTTCAAGGGTATAATTATTTTAAGCAACTC
ACTTGGAAAGAATCATGACTTCTGTCACTGCTTTTATCCTTGTTGCAGTTCTTTTAATAC
AGGGCCCTTTGAGAGCCCATTAATTTATGACTGGGTTTCTCGTTACTTTGAGTAAATTAG
AGTGTTTAAAGCAAACAGATAAAGCGTATTTTACTGTGTTTGAATACTATAGCATGGAAT
AACAACATTGAATAGGTCAAAAGTTTTTGAAAAATTTTTCTTATTTTGGCTTAGATACAG
TTAATAGGAGTAGCTTGGGGGCATTTGTATTCAGATGTCAGAGGTGAAATTCTTAGATTT
TCTGGAGACAAACAACTGCGAAAGCATTTGCCTAAAATACTTCCATTAATCAAGAACGAA
AGTTAAGGGAGTGAAGACGATCAGATACCGTCGTAATCTTAACCATAAACTATGCCGACT
AAGTGTTGGATGAAAATTTATAAATAAAACTATCTTCTTTAAAGGAGTAGTTTTTTAGAT
GCTTCCTTCAGTACCTTATGAGAAATCAAAGTCTTTGGGTTCTGGGGCGAGTATTCGCGC
AAGCGAGAAAGTTAAAAGAATTGACGGAAGGGCACCACCAGGCGTGGAGCTTGCGGCTTA
ATTTGACTCAACACGGGGAAACTCACTAGTTTAAGACAAGAGTAGGATTGACAGATTAAT
AGCTCTTTCTTGATTTCTTGGATGGTGATGCATGGCCGTTTTTAGTTCGTGAATATGATT
TGTCTGGTTAATTCCGATAACGAACGAGATCTTAACCTGCTAATTAGCGGTGGATATGTG
ATATTCTTCGAAGGTGGACTAACTATAGCGTTTTCGAAGGTATGTTGCATAATCAAATTG
GTTTACCCTTTGTTTTTTTGTAGCATATTCTTTTATTTCGTTGGGTTTTTTCCCTAGTAA
GGATGTATCTGCTTTATTTAATGCTTCTTAGAGGAACGATGTGTGTCTAACACAAGGAAG
TTTAAGGCAACAACAGGTCTGTGATGTCCTTAGATATACTAGGCTGCACGCGTGCTACAC
TGATATGTAAAACGAGTATTTAAAATTATATCTGTATGGTAGATAATTTAATTTCTACGT
ATTATCAGCATATACTTTTCCTACACTGAAATAGTGAAGGTAATCTTTATCAATACATAT
CGTGATGGGGATAGATTATTGCAATTATTAATCTTGAACGAGGAATGCCTAGTAAGCATG
ATTCATCAGATTGTGCTGACTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTCCTAC
CGATTGAAAGATATGATGAATTGTTTGGACAAGAAAATAGAAATTTTATTTTTATTTTTT
TTGGAAGGACCGTAAATCCTATCTTTTAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTA
GGTGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCA
ACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCG
CACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTA
TTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAAT
CTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCC
CTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAG
CTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGT
GATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGG
CACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAA
TATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA
GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCT
TCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGG
TGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCG
CCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATT
ATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGA
CTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGA
ATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAAC
GATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCG
CCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCAC
GATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCT
AGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCT
GCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGG
GTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTAT
CTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGG
TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGAT
TGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCT
CATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAA
GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAA
AAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCC
GAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTA
GTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCT
GTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACG
ATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAG
CTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGC
CACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGG
AGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTT
TCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATG
GAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCA
CATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTG
AGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGC
GGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAG
CTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAG
TTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTG
TGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAA
GCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACA
TATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAA
ATATATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTT
GTATGTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGA
CACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATAT
TTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTA
TTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAAT
AATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAA
TACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATATATATA
TATATATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAAT
AATATATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCAT
ACATGCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTA
TGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTT
TATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATT
TTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGA
ARGTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTT
ATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCA
ATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAA
GTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTAC
ACATTAAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAGGGAAAT
TCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGA
TATATACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGT
ATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCT
GTTTTTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATA
TAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCY
TGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAA
ATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTT
TAATATCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTTATATAT
TTATAATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCA
ATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAA
GRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTAT
TTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATAT
AATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGG
TTATATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTA
AGATATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATT
TTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCAT
TTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTC
ATTTGTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATA
AACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCC
TCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGC
CCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACT
CTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAA
CCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTT
CCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGC
GCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGG
ATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTC
TGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTT
TCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTG
CAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACA
ATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCT
ACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGA
GCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGG
CTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGA
AAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATG
AAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATG
ACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATC
AGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGC
TGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCT
GGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCG
GAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACA
TGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGA
GAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACG
AAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGC
TGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCG
CTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGT
TCATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGC
AGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCA
AGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGAC
GAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTT
CTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTG
TGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTAT
TTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATT
TTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAAC
ATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATA
ATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAA
TATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAA
ATGATGTTTTTTCCTTCAATTTCGAT
Restriction sites to linearize plasmid ApaI/KspI
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe bir38 gene was amplified from genomic DNA, including 1.5 kb of DNA upstream of the ATG of the bir ORF, that abuts to the next gene in the P. berghei genome and was therefore assumed to include the entire promoter.
Amplification of the 2.6 kb BIR38 fragment resulted in loss of the TAA stop condon allowing c-terminal fusion of eGFP
Primers used for PCR amplification of bir38: reverse NcoI: CATGCCATGGGATGATTCATTCTCTTC; forward PvuII: TTCCAGCTGGAAACGCGGGAATATTATAC.
The construct contains a fragment of of the non-essential c/d-type ssu-rrna gene unit as a target region for integration into the genome.
The bir38 gene has been integrated into the genome by single cross-over integration. Therefore the possibility exists of reversion to the wild-type genotype by removal of the integrated DNA construct.
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0837161
Gene Model P. falciparum ortholog Not available
Gene productBIR protein PIR protein
Gene product: Alternative nameBIR38, Plasmodium berghei interspersed repeat protein 38
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1TTCCAGCTGGAAACGCGGGAATATTATAC
Additional information primer 1BIR38 + 5'UTR forward (PvuII)
Sequence Primer 2CATGCCATGGGATGATTCATTCTCTTC
Additional information primer 2BIR38 + 5'UTR reverse (NcoI)
3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionInsertion locus
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c-type unit)
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4