RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1442

Malaria parasite	P. yoelii
Genotype
Mutated	Gene model (rodent): PY17X_0405400; Gene model (P.falciparum): PF3D7_0304600; Gene product: circumsporozoite (CS) protein (CS; CSP) Details mutation: The endogenous P. yoelii csp gene has been replaced with the csp gene of P. falciparum.
Phenotype	No phenotype has been described

Last modified: 26 April 2016, 19:10

*RMgm-1442

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene mutation
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 27068454
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. yoelii
Parent strain/line	P. y. yoelii 17XNL
Name parent line/clone	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Zhang M; Yuda, M; Tsuij M
Name Group/Department	HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center
Name Institute	Affiliate of The Rockefeller University
City	New York
Country	USA
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Name of the mutant parasite
RMgm number	RMgm-1442
Principal name	PfCSP/Py
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	Not different from wild type
Liver stage	Not different from wild type
Additional remarks phenotype	Mutant/mutation In the mutant the endogenous P. yoelii csp gene has been replaced with the csp gene of P. falciparum. The P. falciparum csp gene is under the control of the promoeter region of P. falciparum csp and the 3'UTR region of hsp70. Protein (function) Phenotype The mutant produced normal numbers of infectious sporozoites Additional information No information is provided on the source (strain, sequence) of the P. falciparum csp sequence Other mutants

Mutated: Mutant parasite with a mutated gene

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Details of the target gene

Gene Model of Rodent Parasite

PY17X_0405400

Gene Model P. falciparum ortholog

PF3D7_0304600

Gene product

circumsporozoite (CS) protein

Gene product: Alternative name

CS; CSP

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Details of the genetic modification

Short description of the mutation

The endogenous P. yoelii csp gene has been replaced with the csp gene of P. falciparum.

Inducable system used

Short description of the conditional mutagenesis

Not available

Additional remarks inducable system

Type of plasmid/construct

(Linear) plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used

Modified PlasmoGEM construct/vector used

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

Additional remarks genetic modification

Transgenic parasites were generated by inserting the PfCSP expression construct at the locus of the P. yoelii CSP gene by double cross-over homologous recombination. The targeting plasmid for generating PfCSP/Py parasites was generated in a pBluescript plasmid that contains the PyCSP promoter, PfCSP coding sequence, 3′UTR of HSP70, human DHFR expression cassette, and 3′UTR of PyCSP. The plasmid was linearized by digestion with restriction enzymes KpnI and NotI.

Additional remarks selection procedure

Primer information: Primers used for amplification of the target sequences Click to view information

Primer information: 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
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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