RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1447

Malaria parasite	P. berghei
Genotype
Mutated	Gene model (rodent): PBANKA_0201800; Gene model (P.falciparum): PF3D7_0112000; Gene product: TatD-like deoxyribonuclease (TatD) Details mutation: an aspartic acid-to-alanine mutation at the 352 position of PbTatD
Phenotype	Asexual bloodstage;

Last modified: 15 May 2016, 21:59

*RMgm-1447

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) : 27151551
MR4 number
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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	Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Chang Z; Chen Q
Name Group/Department	Key Laboratory of Zoonosis
Name Institute	Jilin University
City	Changchun
Country	China
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Name of the mutant parasite
RMgm number	RMgm-1447
Principal name	PbTatD-D352A
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	(Slightly) reduced growth rate of blood stages in vivo.
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expresses a mutated form of TatD (an aspartic acid-to-alanine mutation at the 352 position of PbTatD). Protein (function) The predicted P. falciparum TatD 3D model (without its signal peptide) contains a hydrolase active site surrounded by seven a-helices. The PfTatD domain contains a TIM barrel (COG 1099, multiple domain), which represents a nucleic acid binding region. Furthermore, compared to E. coli TatD, Plasmodium TatD contains an insertion in the middle of the sequence consisting of 73 amino acids (AAs), which marks another feature of the Plasmodium TatDs that is distinct from other microbial homologues, apart from the signal peptide. Phenotype (Slightly) reduced growth rate of blood stages in vivo (and prolonged survial of mice). See also RMgm-1446 for a mutant lacking expression of TatD. This mutant has a similar phenotype. Additional information Evidence is presented that TatD is exported into the cytoplasm of infected red blood cells. Evidence is presented for deoxyribonuclease/DNA hydrolase activity. Other mutants See also RMgm-1446 for a mutant lacking expression of TatD. This mutant has a similar phenotype.

Mutated: Mutant parasite with a mutated gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0201800

Gene Model P. falciparum ortholog

PF3D7_0112000

Gene product

TatD-like deoxyribonuclease

Gene product: Alternative name

TatD

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Details of the genetic modification

Short description of the mutation

an aspartic acid-to-alanine mutation at the 352 position of PbTatD

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

pbdhfr

Promoter of the selectable marker

pbdhfr

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

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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