JOAO EZEQUIEL DE OLIVEIRA

JOAO EZEQUIEL DE OLIVEIRA

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Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs
2017 - SEVILHANO, THAIS; CARVALHO, ROBERTO F. de; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E.; MALTAROLLO, VINICIUS G.; TROSSINI, GUSTAVO; GARCEZ, RIVIANE; BARTOLINI, PAOLO
The common gonadotrophic hormone α-subunit (GTHα) has been previously isolated by our research group from A. gigas pituitaries; in the present work the cDNA sequences encoding FSHβ and LHβ subunits have also been isolated from the same species of fish. The FSH β-subunit consists of 126 amino acids with a putative 18 amino acid signal peptide and a 108 amino acid mature peptide, while the LH β-subunit consists of 141 amino acids with a putative 24 amino acid amino acid signal peptide and a 117 amino acid mature peptide. The highest identity, based on the amino acid sequences, was found with the order of Anguilliformes (61%) for FSHβ and of Cypriniformes (76%) for LHβ, followed by Siluriformes, 53% for FSHβ and 75% for LHβ. Interestingly, the identity with the corresponding human amino acid sequences was still remarkable: 45.1% for FSHβ and 51.4% for LHβ. Three dimensional models of ag-FSH and ag-LH, generated by using the crystal structures of h-FSH and h-LH as the respective templates and carried out via comparative modeling and molecular dynamics simulations, suggested the presence of the so-called ªseat-beltº, favored by a disulfide bond formed between the 3rd and 12th cysteine in both β-subunits. The sequences found will be used for the biotechnological synthesis of A. gigas gonadotrophic hormones (ag-FSH and ag-LH). In a first approach, to ascertain that the cloned transcripts allow the expression of the heterodimeric hormones, ag-FSH has been synthesized in human embryonic kidney 293 (HEK293) cells, preliminarily purified and characterized.
The influence of Escherichia coli cultivation temperature on interferon alpha 2a expression (IFN-'alfa'2a)
2014 - ARTHUSO, FERNANDA dos S.; SUZUKI, MIRIAM F.; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E. de; BARTOLINI, PAOLO; SOARES, CARLOS R.J.
Two-step purification of mouse prolactin expressed in the periplasmic space of Escherichia coli
2010 - SUZUKI, MIRIAM F.; ARTHUSO, FERNANDA dos S.; SOUZA, JOSE M.; OLIVEIRA, JOAO E.; OLIVEIRA, TAIS L.; OLIVEIRA, NELIO A. de J.; GOULART, HERBERT R.; CAPONE, MARCOS V.N.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.
The influence of Escherichia coli cultivation temperature on interferon alpha expression (IFN-'alfa'2a)
2013 - ARTHUSO, FERNANDA dos S.; SUZUKI, MIRIAM F.; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E. de; BARTOLINI, PAOLO; SOARES, CARLOS R.J.
Expressao de prolactina de camundongos no espaco periplastico de Escherichia coli
2009 - SUZUKI, MIRIAM F.; ARTHUSO, FERNANDA dos S.; SOUZA, JOSE M.; OLIVEIRA, JOAO E.; OLIVEIRA, TAIS L.; OLIVEIRA, NELIO A. de J.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.