1988
DOI: 10.4049/jimmunol.141.10.3584
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Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii.

Abstract: The complete sequence of P30, the major surface Ag of the protozoan parasite, Toxoplasma gondii, has been deduced through the cloning and analysis of its gene. Using polyclonal serum specific for P30, we have isolated a P30 cDNA clone from a lambda gt11 cDNA expression library derived from tachyzoites of T. gondii (RH strain). This clone produces a beta-galactosidase fusion protein which reacts with several anti-P30 mAb. In addition, polyclonal anti-serum raised to the fusion protein reacts with purified P30 p… Show more

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Cited by 378 publications
(22 citation statements)
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“…Parasites were settled for 10 min at RT and incubated at 37°C for 20 min, followed by fixation of 4% PFA for 15 min. Sample was washed 3× with PBS, blocked without Triton (3% BSA in PBS) for 1 h, stained with anti-Sag1 antibody [ 66 ], washed 3× with PBS, and stained with anti-mouse Alexa-488. Sample was briefly fixed with PFA, washed 3× with PBS, blocked with Triton (3% BSA in PBS, 0.2% Triton-X-100) for 1 h, stained with α-Gap45 antibody, washed 3× with PBS, and stained with anti-rabbit Abberior-580.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Parasites were settled for 10 min at RT and incubated at 37°C for 20 min, followed by fixation of 4% PFA for 15 min. Sample was washed 3× with PBS, blocked without Triton (3% BSA in PBS) for 1 h, stained with anti-Sag1 antibody [ 66 ], washed 3× with PBS, and stained with anti-mouse Alexa-488. Sample was briefly fixed with PFA, washed 3× with PBS, blocked with Triton (3% BSA in PBS, 0.2% Triton-X-100) for 1 h, stained with α-Gap45 antibody, washed 3× with PBS, and stained with anti-rabbit Abberior-580.…”
Section: Methodsmentioning
confidence: 99%
“…Per replica and condition 250 μl of parasites were added on a 24-well plate with confluent HFF on glass slides, allowed to settle for 20 min at RT, and then incubated for 20 min at 37°C. Cells were fixed with ice-cold methanol at −20°C for 8 min and stained with anti-STAT6-P (1:600, (Cell Signaling 56554S)) and anti-Sag1 antibody [ 66 ].…”
Section: Methodsmentioning
confidence: 99%
“…Tachyzoites treated with IAA or vehicle for three growth cycles were inoculated onto coverslips with confluent monolayers of HFFs and grown for 24 h. Cells were fixed in 4% (vol/vol) paraformaldehyde/ PBS (Sigma-Aldrich) for 10 min and permeabilised in 0.1% (vol/vol) Triton X-100/PBS before blocking in 3% (wt/vol) BSA/PBS (Sigma-Aldrich) for 1 h. Cells were incubated with primary antibodies used at 1:1,000 in 3% BSA/PBS for 1 h. Primary antibodies used were as follows: rat α-HA 3F10 (1:1,000, Cat# 11867423001, RRID:AB_390918; Roche); mouse α-SAG1 DG52 (Burg et al, 1988); rabbit α-GAP45 (1:1,000) (Gaskins et al, 2004); rabbit α-ACP (Waller et al, 1998); and rabbit α-IMC1 (Ward & Carey, 1999) (Table S7). Cells were washed 3X in PBS and probed for 1 h with Alexa Fluor-conjugated secondary antibodies (Thermo Fisher Scientific; see Table S8 for full list), plus 5 μg/ml DAPI used at 1:1,000 in 3% BSA/PBS.…”
Section: If Microscopymentioning
confidence: 99%
“…Of note, these strains all constitutively express an RFP (WT III expresses mCherry and the others express tdTomato). We induced encystment by placing the infected cultures under alkaline stress (pH 8.2) in combination with CO 2 deprivation (<5%) and tracked differentiation using immunofluorescent staining of T. gondii stage-specific surface antigens [41,42] and Dolichos biflorus agglutinin (DBA), which binds sugar moieties present on the proteins that form the cyst wall [43]. At 6 days post infection (dpi) the WT III strain no longer expressed the tachyzoite-specific antigen SAG1 [41], instead expressing the bradyzoite-specific surface antigen SRS9 [42] and staining positive for DBA, indicating formation of the cyst wall (S1 Fig) . In contrast, IIIΔrop16 parasites displayed a defect in SRS9 expression and DBA staining, which could be rescued by complementation with ROP16 III but not by complementation with ROP16 II (S1 Fig).…”
Section: Rop16 Facilitates Cyst Development In a Strain-specific Mannermentioning
confidence: 99%