Cloning and expression of a chimera containing ROP2 of Neospora caninum fused with OprI lipoprotein from Pseudomonas aeruginosa / Clonagem e expressão de quimera contendo a proteína ROP2 de Neospora caninum fusionada a lipoproteína OprI de Pseudomonas aeruginosa

Neospora caninum is the etiologic agent of neosporosis, is one of the main responsible for abortion in cattle herds, causing economic losses to Cattle-raising. Vaccination of cattle would be an important alternative; however, the lack of effective vaccines prevents the application of this control method. The proteins present in rhoptries (ROPs), due to their importance in cell infection and their antigenic and immunogenic characteristics, became excellent candidates for vaccine antigens. The bacterial lipoproteins as an Oprl from Pseudomonas aeruginosa have received particular attention as an adjuvant carrier molecule. The aims of this study were to clone and expressing a chimera containing NcROP2 fused with OprI lipoprotein from P. aeruginosa for the future development of a recombinant vaccine against N. caninum. We cloned and expressed, in Escherichia coli Rosetta (DE3) pLysS, the region of NcROP2 described between amino acids 191 and 359, fused OprI producing the chimera rROP2/OprI, showed an expected size of ~50 kDa. and characterized its antigenicity. The protein was purified and characterized by Western blot with anti-histidine monoclonal antibodies and their antigenicity recognized by sera from animals naturally infected by N. caninum. The chimera rROP2/OprI was recognized by antibodies anti-N. caninum reviling common antigenic determinants of the recombinant protein and its native form, suggesting its use for developing a recombinant vaccine

Zika virus disrupts molecular fingerprinting of human neurospheres

Submitted by Sandra Infurna ([email protected]) on 2018-04-03T16:26:16Z No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-04-03T16:44:04Z (GMT) No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5)Made available in DSpace on 2018-04-03T16:44:04Z (GMT). No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5) Previous issue date: 2017Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Ciências Biológicas. Rio de Janeiro, RJ, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto Evandro Chagas. Centro de Inovação Tecnológica. Belém, PA, BrasilInstituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilInstituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Fundação Oswaldo Cruz Fiocruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade Federal do Pará. Belém, PA, Brasil.Fundação Oswaldo Cruz Fiocruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Instituto Evandro Chagas. Centro de Inovação Tecnológica. Belém, PA, BrasilUniversidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Ciências Biológicas. Rio de Janeiro, RJ, Brasil.Zika virus (ZIKV) has been associated with microcephaly and other brain abnormalities; however, the molecular consequences of ZIKV to human brain development are still not fully understood. Here we describe alterations in human neurospheres derived from induced pluripotent stem (iPS) cells infected with the strain of Zika virus that is circulating in Brazil. Combining proteomics and mRNA transcriptional profiling, over 500 proteins and genes associated with the Brazilian ZIKV infection were found to be differentially expressed. These genes and proteins provide an interactome map, which indicates that ZIKV controls the expression of RNA processing bodies, miRNA biogenesis and splicing factors required for self-replication. It also suggests that impairments in the molecular pathways underpinning cell cycle and neuronal differentiation are caused by ZIKV. These results point to biological mechanisms implicated in brain malformations, which are important to further the understanding of ZIKV infection and can be exploited as therapeutic potential targets to mitigate it