Comparative study between the efficiency of transfection and co-transfection of vectors carrying the gene of the rabies virus glycoprotein (GPV) in cells of Drosophila melanogaster. / Estudo comparativo entre a eficiência de co-transfecção e transfecção de vetores portadores do gene da glicoproteína do vírus rábico (GPV) em células de Drosophila melanogaster S2.

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

Among the advantages of the system of gene expression in cells drosófila there is still the rapid establishment of stable cell lines that secrete efficiently the recombinant protein. We have established populations of cells co-transfected S2AcGPV and transfected S2AcGPVHy. The objective of this study is a comparison of the expression of GPV in S2 cells co-transfected with vector pAcGPV (vector of expression) and pCoHygro (vector of selection) or transfected with a single vector pAcGPVHygro (containing the vector of expression and selection gene). The populations were analyzed in relation to the expression of GPV in immunoassays: ELISA test, Dot Blot, the SDS-PAGE gels, Western Blot, flow cytometry (FACS) and confocal microscopy. Tests of immunofluorescence in flow cytometry (FACS) have shown that cells co-transfected and transfected are expressing the protein GPV. Values between 0.3 and 4 mg/107células were obtained. Moreover, anti-GPV were able to recognize the protein GPV.

ASSUNTO(S)

biologia molecular drosophila melanogaster biotecnologia expressão gênica biotechnology cells transfection transfecção molecular biology células gene expression insects insetos drosophila melanogaster

ACESSO AO ARTIGO

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