Data/hora: 08/06/2024 08:22:50

Provedor de dados: | Brazilian Journal of Medical and Biological Research
País: | Brazil
Título: | Expression of the Mycobacterium bovis P36 gene in Mycobacterium smegmatis and the baculovirus/insect cell system
Autores: | Bigi,F.; Taboga,O.; Romano,M.I.; Alito,A.; Fisanotti,J.C.; Cataldi,A.A.
Data: | 1999-01-01
Ano: | 1999
Palavras-chave: | Mycobacterium tuberculosis; Mycobacterium smegmatis; Insect cells; Baculovirus; Secreted proteins; P36; MPB70; Antigen.
Resumo: | In the present study we evaluated different systems for the expression of mycobacterial antigen P36 secreted by Mycobacterium bovis. P36 was detected by Western blot using a specific antiserum. The P36 gene was initially expressed in E. coli, under the control of the T7 promoter, but severe proteolysis prevented its purification. We then tried to express P36 in M. smegmatis and insect cells. For M. smegmatis, we used three different plasmid vectors differing in copy number and in the presence of a promoter for expression of heterologous proteins. P36 was detected in the cell extract and culture supernatant in both expression systems and was recognized by sera from M. bovis-infected cattle. To compare the expression level and compartmentalization, the MPB70 antigen was also expressed. The highest production was reached in insect cell supernatants. In conclusion, M. smegmatis and especially the baculovirus expression system are good choices for the production of proteins from pathogenic mycobacteria for the development of mycobacterial vaccines and diagnostic reagents.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000100004
Editor: | Associação Brasileira de Divulgação Científica
Relação: | 10.1590/S0100-879X1999000100004
Formato: | text/html
Fonte: | Brazilian Journal of Medical and Biological Research  v.32 n.1 1999
Fonte: | info:eu-repo/semantics/openAccess