Visualization of a Recombinant Gene Protein in the Baculovirus Expression Vector System Using Confocal Scanning Laser Microscopy

概要

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Expression of the recombinant protein β-galactosidase in the Spodoptera frugiperda Sf-9 insect cell line infected by the Autographa californica nuclear polyhedrosis virus expressing β-galactosidase(AcNPV-βgal)was visualized using confocal scanning laser microscopy with fluorescent staining of both the recombinant protein and the cell nucleus. The average size of the insect cells and the intracellular DNA concentration both increased markedly, respectively reading 3.8- and 2.3-fold the values before infection. The average β-galactosidase activity began to increase at 20-24h post infection and finally reached 1.9×10^4 units/ml. As the post infection time increased, the stained nucleus images expanded and spread broadly. β-Galactosidase was first identified by fluorescent staining at 12h post-infection, filled the cell at 27h, began to be released at 36h, and finally spread out of the cell. The locations of the nucleus and expressed β-galactosidase were identified from computerized tomograms and 3-dimensional images.
社団法人日本生物工学会の論文
1999-06-25