Expression of the hepatitis B virus surface antigen in Drosophila S2 cells

Autor(es)

Jorge‚ S.A.C. ; Santos‚ A.S. ; Spina‚ Â. ; Pereira‚ C.A.

• Tipo de documento: Artigo Científico
• Fonte: Cytotechnology; v. 57, n. 1, p. 51-59, 2008.

Resumo

Drosophila melanogaster S2 cells were transfected with a plasmid vector (pAcHBsAgHy) containing the S gene, coding for the hepatitis B virus surface antigen (HBsAg), under control of the constitutive drosophila actin promoter (pAc), and the hygromycin B (Hy) selection gene. The vector was introduced into Schneider 2 (S2) Drosophila cells by DNA transfection and a cell population (S2AcHBsAgHy) was selected by its resistance to hygromycin B. The pAcHBsAgHy vector integrated in transfected S2 cell genome and approximately 1,000 copies per cell were found in a higher HBsAg producer cell subpopulation. The HBsAg production varied in different subpopulations, but did not when a given subpopulation was cultivated in different culture flasks. Higher HBsAg expression was found in S2AcHBsAgHy cells cultivated in Insect Xpress medium (13.5 mu g/1E7 cells) and SFX medium (7 mu g/1E7 cells) in comparison to SF900II medium (0.6 mu g/1E7 cells). An increase of HBsAg was observed in culture maintained under hygromycin selection pressure. Data presented in the paper show that S2AcHBsAgHy cells produce efficiently the HBsAg which is mainly found in the cell supernatant, suggesting that HBsAg is secreted from the cells. The data also show that our approach using the Drosophila expression system is suitable for the preparation of other viral protein preparation. (AU)

• Processo FAPESP: 05/50565-8 - Construcao de vetores e estrategias para expressao dos genes do virus da hepatite b (hbsag) e do virus da raiva (gpv/pv) em celulas de drosofila.
• Beneficiário: Carlos Augusto Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 02/09482-3 - Expressão de genes heterólogos em células de dípteros: biologia molecular e engenharia de processos
• Beneficiário: Carlos Augusto Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 01/08914-4 - Expressão dos genes do vírus da hepatite B (HBsAg) e do vírus da raiva (GPV/PV) em células de drosófilas
• Beneficiário: Soraia Attie Calil Jorge
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado