Molecular cloning, sequencing and expression of a serine proteinase inhibitor gene from Toxoplasma gondii

dc.creatorPszenny, Viviana
dc.creatorAngel, Sergio O.
dc.creatorDuschak, Vilma G
dc.creatorPaulino, M
dc.creatorLedesma, Bibiana A
dc.creatorYabo, M I
dc.creatorGuarnera, Eduardo
dc.creatorRuiz, Andrés M
dc.creatorBontempi, Esteban
dc.date2019-12-12T20:30:23Z
dc.date2019-12-12T20:30:23Z
dc.date2000-04-15
dc.date.accessioned2023-08-29T20:06:40Z
dc.date.available2023-08-29T20:06:40Z
dc.identifier0166-6851
dc.identifierhttp://sgc.anlis.gob.ar/handle/123456789/1501
dc.identifier10.1016/s0166-6851(00)00202-4
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8519241
dc.descriptionFil: Pszenny, Viviana. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionFil: Angel, Sergio O. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionFil: Duschak, Vilma G. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina.
dc.descriptionFil: Paulino, Margot. Universidad de la República. Facultad de Química. Cátedra de Química Cuántica, Montevideo; Uruguay.
dc.descriptionFil: Ledesma, Bibiana A. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionFil: Yabo, Miriam I. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Producción de Biológicos; Argentina.
dc.descriptionFil: Guarnera, Eduardo. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionFil: Ruiz, Andrés M. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionFil: Bontempi, Esteban. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Parasitología; Argentina.
dc.descriptionA cDNA clone from a Toxoplasma gondii tachyzoite cDNA library encoding a serine proteinase inhibitor (serpin) was isolated. The 1376 bp cDNA sequence encodes a 294 amino acid protein with a putative signal peptide of 23 amino acids resulting in a mature protein with a predicted mass of 30,190 Da and a pI of 4.86. This protein has internal sequence similarity of residues 30-66, 114-150, 181-217 and 247-283 indicating a four-domain structure. The four domains exhibit high identity to serine proteinase inhibitors belonging to the non-classical Kazal-type family. The gene is single copy in the tachyzoite haploid genome of RH strain and was amplified by polymerase chain reaction (PCR). Several introns were identified. The sequence encoding the mature protein was amplified by PCR, cloned into the pQE30 vector and expressed in Escherichia coli. Specific antiserum generated against the recombinant protein was used in immunoblot assay and two bands of 38 and 42 kDa were detected in a whole parasite homogenate. The recombinant protein showed trypsin-inhibitory activity, one of the two potential specificities. We discuss the possible roles that T. gondii serpin(s) may play in the survival of the tachyzoites in the host.
dc.formatpdf
dc.languageen
dc.relationMolecular and biochemical parasitology
dc.rightsnone
dc.sourceMolecular and Biochemical Parasitology 2000;107(2):241-249
dc.subjectInhibidor de Serinpeptidasas Tipo Kazal-5
dc.subjectInhibidores de Serinpeptidasas Tipo Kazal
dc.subjectToxoplasma
dc.titleMolecular cloning, sequencing and expression of a serine proteinase inhibitor gene from Toxoplasma gondii
dc.typeArtículo


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