Comparative Proteomic Analysis of Toxoplasma gondii RH Wild-Type and Four SRS29B (SAG1) Knock-Out Clones Reveals Significant Differences between Individual Strains
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Host Cell Culture, Maintenance, and Purification of Tachyzoites
4.3. Proteomics
4.4. Quantification of Transcripts by Multiplex TaqMan—qPCR
4.5. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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GN | Annotation | WT/C6 | WT/C18 | WT/C30 | WT/C33 |
---|---|---|---|---|---|
TGRH88_003360 | Unspecified product (putative transmembrane protein) | down | down | ||
TGRH88_006930 | SAG-related sequence srs16C | up | |||
TGRH88_012840 | Unspecified product (putative transmembrane protein) | up | |||
TGRH88_016110 | SAG-related sequence srs36D | up | up | up | |
TGRH88_017250 | Unspecified product | up | up | up | |
TGRH88_021190 | cAMP-dependent protein kinase | down | |||
TGRH88_029080 | Gamma-glutamyl hydrolase | up | |||
TGRH88_029100 | Unspecified product | down | |||
TGRH88_036500 | Multi-pass transmembrane protein | down | down | ||
TGRH88_042260 | Unspecified product (putative transmembrane protein) | up | up | ||
TGRH88_043760 | Putative translation elongation and release factors (GTPase) | down | down | ||
TGRH88_046380 | Histone lysine methyltransferase set1 | down | down | ||
TGRH88_055240 | Putative myosin heavy chain | up | |||
TGRH88_062920 | Bifunctional dihydrofolate reductase—thymidylate synthase | down | down | ||
TGRH88_073870 | MaoC family domain-containing protein | up | |||
TGRH88_077450 | SAG-related sequence srs29B | up | up | up | up |
GN | Annotation | C6/C18 | C6/C30 | C6/C33 | C18/C30 | C18/C33 | C30/C33 |
---|---|---|---|---|---|---|---|
TGRH88_004120 | PRELI family protein | up | |||||
TGRH88_006930 | SAG-related sequence srs16C | up | up | ||||
TGRH88_008350 | Unspecified product (putative iron-sulfur cluster assembly accessory protein) | down | down | ||||
TGRH88_009830 | SAG-related sequence srs19F | down | |||||
TGRH88_012840 | Unspecified product (Putative transmembrane protein) | down | down | ||||
TGRH88_014370 | Cathepsin cpc1 | down | up | ||||
TGRH88_016110 | SAG-related sequence srs36D | down | down | down | up | ||
TGRH88_016870 | Unspecified product | up | up | ||||
TGRH88_017250 | Unspecified product | down | down | ||||
TGRH88_021190 | cAMP-dependent protein kinase | up | up | up | |||
TGRH88_046380 | Histone lysine methyltransferase set1 | down | down | ||||
TGRH88_076630 | Unspecified product | down |
Gene ID (ToxoDB) | Gene Name | Forward Primer (5′-3′) | Reverse Primer (5′-3′) | Taqman-Hydrolysis Probe |
---|---|---|---|---|
TGRH88_014440 | Glyceraldehyde-3-phosphate dehydro-genase GAPDH1 | TGGAGGTTTTGGCGAT | ATGGCAGTTGGCTCCTT | HEX-TACCCCGGC-GAAGTCAGC-BHQ |
TGRH88_016080 | SAG-related sequence SRS36B (SAG 5D) | TCAGAGGTCACCCGAGT | TCGTGGTGTCCTGGTTAC | FAM-CACCATCCAG-GTGATCCAGCC-BHQ |
TGRH88_016110 | SAG-related sequence SRS36D (SAG 5C) | TGTATGGCAAGCGACAA | TGTGCACACCTCAGTTAGTG | FAM-ACGGGCAG-AGTGACAGCGCA-BHQ |
TGRH88_040330 | SAG-related sequence SRS46 | TGCAGACGTACCGACAGT | TCTGCGTCGACGAGTG | FAM-TCTTCGCTG-CGGCAAAAACTT-BHQ |
TGRH88_055140 | Alpha tubulin TUBA1 | GACGCCTTCAACACCTTC | TTGTTCGCAGCATCCT | Cy5-TTACCGCCA-CCTGTTCCACCC-BHQ |
TGRH88_057630 | SAG-related sequence SRS57 (SAG 3) | TGCGATCTTGGGAAC | TCAGGGTGCTCTTTGTTC | FAM-TGTTCGTCG-CCGCAGGGA-BHQ |
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Hänggeli, K.P.A.; Hemphill, A.; Müller, N.; Heller, M.; Uldry, A.-C.; Braga-Lagache, S.; Müller, J.; Boubaker, G. Comparative Proteomic Analysis of Toxoplasma gondii RH Wild-Type and Four SRS29B (SAG1) Knock-Out Clones Reveals Significant Differences between Individual Strains. Int. J. Mol. Sci. 2023, 24, 10454. https://doi.org/10.3390/ijms241310454
Hänggeli KPA, Hemphill A, Müller N, Heller M, Uldry A-C, Braga-Lagache S, Müller J, Boubaker G. Comparative Proteomic Analysis of Toxoplasma gondii RH Wild-Type and Four SRS29B (SAG1) Knock-Out Clones Reveals Significant Differences between Individual Strains. International Journal of Molecular Sciences. 2023; 24(13):10454. https://doi.org/10.3390/ijms241310454
Chicago/Turabian StyleHänggeli, Kai Pascal Alexander, Andrew Hemphill, Norbert Müller, Manfred Heller, Anne-Christine Uldry, Sophie Braga-Lagache, Joachim Müller, and Ghalia Boubaker. 2023. "Comparative Proteomic Analysis of Toxoplasma gondii RH Wild-Type and Four SRS29B (SAG1) Knock-Out Clones Reveals Significant Differences between Individual Strains" International Journal of Molecular Sciences 24, no. 13: 10454. https://doi.org/10.3390/ijms241310454