RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-803

Malaria parasite	P. yoelii
Genotype
Tagged	Gene model (rodent): PY17X_0303100; Gene model (P.falciparum): PF3D7_0204700; Gene product: hexose transporter (hexose transporter 1, HT1; PfHT1, PfHT, PbHT, PbHT1) Name tag: HA tag
Phenotype	Sporozoite;

Last modified: 19 January 2013, 14:18

*RMgm-803

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 23325771
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	S.E. Lindner; S.H. Kappe
Name Group/Department	Malaria Program
Name Institute	Seattle Biomedical Research Institute
City	Seattle
Country	US
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Name of the mutant parasite
RMgm number	RMgm-803
Principal name	PY00899ha
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Not tested
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	HA-tagged expression of PY00899 was detected in salivary gland sporozoites. A strong circumferential staining of the sporozoite was obtained with antibodies to the HA tag that co-localized well with CSP
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a C-terminal HA-tagged version of PY00899 Protein (function) The protein was detected in a 'sporozoite surface-exposed' proteome analysis of sporozoites. Phenotype Expression was only analysed in the sporozoite stage. HA-tagged expression of PY00899 was detected in salivary gland sporozoites. A strong circumferential staining of the sporozoite was obtained with antibodies to the HA tag that co-localized well with CSP, indicating a surface location. See also P. berghei mutants RMgm-391 and RMgm-603 that express tagged-versions of this hexose transporter Additional information In this study putative surface-exposed proteins from P. falciparum and P. yoelii salivary gland sporozoites were identified by specifically labeling live sporozoites with a biotin-conjugated, amine-reactive cross-linker and subsequent pull-down by streptavidin-coated Dynabeads. Other mutants See P. berghei mutants RMgm-391 and RMgm-603 that express tagged-versions of this hexose transporter

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PY17X_0303100

Gene Model P. falciparum ortholog

PF3D7_0204700

Gene product

hexose transporter

Gene product: Alternative name

hexose transporter 1, HT1; PfHT1, PfHT, PbHT, PbHT1

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Details of the genetic modification

Name of the tag

HA tag

Details of tagging

C-terminal

Additional remarks: tagging

a triple HA epitope tag on the C-terminus of the PY00899 protein

Commercial source of tag-antibodies

Type of plasmid/construct

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

not reported

Promoter of the selectable marker

not reported

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

No information is provided on the DNA primers and construct used for tagging PY00899

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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