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Frammenti antigenici di Toxoplasma gondii, metodo per il loro ottenimento, loro uso in kit diagnostici e per la preparazione di vaccini
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Studio dei geni per le proteine ribosomali in Xenopus laevis: struttura e meccanismi di regolazione al livello traduzionale
No full textThe SAG5 genes of Toxoplasma gondii are differentially expressed in tachyzoites and bradyzoites and in parasite strains of distinct virulence
No full textInteraction of proteins with the mRNA for ribosomal protein L1 in Xenopus: structural characterization of in vivo complexes and identification of proteins that bind in vitro to its 5'UTR
No full textXenopus r-protein mRNAs are known to be coordinately regulated at the translational level. To find out if RNA/protein interactions are involved in this control mechanism, we have characterized the particles containing the translationally repressed rp-mRNA and we have investigated the proteins that specifically bind to this type of mRNA. By sedimentation analysis and isopycnic centrifugation we have found that the repressed rp-mRNAs are assembled in slow sedimenting complexes where the RNA is prevalent over the protein mass (2.3 to 1). This composition is maintained also after in vitro reconstitution of the particle. We carried out also a detailed analysis of in vitro RNA/protein complex formation by focusing our attention on the 5'UTR, very similar in different rp-mRNAs and important in the translational regulation. We describe specific interactions of L1 mRNA with four proteins. The binding site of two of them, 57 kD and 47 kD, is in the typical pyrimidine sequence at the 5' end and is position dependent. Proteins of the same size interact also with the analogous region of r-protein S1 and L14 mRNA, not with unrelated RNAs. Binding of two other proteins, 31 kD and 24 kD, in the downstream region of the 5'UTR was also observed. The most evident 57 kD protein has been partially purified. Although the binding of these proteins to the r-protein mRNA 5'UTR is specific, their involvement in the translation regulation remains to be proved
Caratterizzazione della struttura e dell'espressione del gene Homeobox Athb-10 di Arabidopsis thaliana
No full textSviluppo di una piattaforma microarray per la diagnosi selettiva dei sottotipi del virus dell'influenza
No full textThrombospondin" and "Sag" protein families of Cryptosporidium parvum and Toxoplasma gondii: cloning and characterization of new molecules potentially involved in parasite-host cell interaction
No full textGene discovery and genetic variation in Cryptosporidium and Toxoplasma. COST820: Vaccines against Animal Coccidioses: “Transfection and whole genome approaches to gene discovery and function: the way forward for vaccine research in coccidians?
No full textAntigen Microarrays for Serodiagnosis of Infectious Diseases
No full textBackground: Progress in robotic printing technology
has allowed the development of high-density nucleic
acid and protein arrays that have increased the throughput
of a variety of assays. We generated protein microarrays
by printing microbial antigens to simultaneously
determine in human sera antibodies directed against
Toxoplasma gondii, rubella virus, cytomegalovirus
(CMV), and herpes simplex virus (HSV) types 1 and 2
(ToRCH antigens).
Methods: The antigens were printed on activated glass
slides with high-speed robotics. The slides were incubated
first with serum samples and subsequently with
fluorescently labeled secondary antibodies. Human IgG
and IgM bound to the printed antigens were detected by
confocal scanning microscopy and quantified with internal
calibration curves. Both microarrays and commercial
ELISAs were utilized to detect serum antibodies
against the ToRCH antigens in a panel of characterized
human sera.
Results: The detection limit (mean 2 SD) of the
microarray assay was 0.5 pg of IgG or IgM bound to the
slides. Within-slide, between-slide, and between-batch
precision profiles showed CVs of 1.7–18% for all antigens.
Overall, >80% concordance was obtained between
microarray assays and ELISAs in the classification of
sera; for T. gondii, CMV, and HSV1, concordance exceeded
90%.
Conclusions: The microarray is a suitable assay format
for the serodiagnosis of infectious diseases and can be
easily optimized for clinical use. The ToRCH assay
performs equivalently to ELISA and may have potentially
important advantages in throughput, convenience,
and cost
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