Expression of human respiratory syncytial virus N and P proteins: functional and immunization studies. / Expressão das proteínas N e P do vírus respiratório sincicial humano: estudos funcionais e de imunização.

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

The Human Respiratory Syncytial Virus is a single stranded negative RNA enveloped virus and it is considered the most important pathogen of the respiratory tract of infants and neonates. The viral N and P proteins were expressed in bacteria, purified and used for the production of polyclonal antibodies in mice. In silico studies allowed the prediction of intrinsically disordered domains for P protein, which were identified by mass spectrometry as protease hyper sensible regions. The optimization of N and P genes allowed a robust expression of the N and P proteins in human cells. DNA vaccines containing the optimized genes were tested in mice and generated strong humoral imune response. The N and P proteins expressed in human cells were immunoprecipitated and their interactions with cellular proteins were identified by mass spectrometry. The P protein was able to interact with the HSP70 protein. Finally, an alternative Minigenome system, using an RNA polymerase II promoter, was developed for HRSV but low or no activity was detected.

ASSUNTO(S)

genetic expression biologia molecular proteins vacinas expressão gênica vaccines bioquímica biochemistry rna viruses vírus de rna molecular biology proteínas

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