Immunocytochemical study of recombinant terminal deoxynucleotidyl transferase (TdT) synthesized by baculovirus-infected insect cells

Abstract

The ability to overproduce terminal transferase through recombinant DNA technology should provide alternate means for generating sufficient quantities for structural and mechanistic study of this creative DNA polymerase. In this work we have investigated, at electron microscope level, the morphological modification and ultrastructural localization of synthesized human terminal transferase occurring in Sf-9 cells during recombinant baculovirus infection time. The results obtained showed that TdT is localized and stored only at the cytoplasmic level; the nucleus did not show any specific site able to link the neosynthesized TdT. The amount of the enzyme, estimate by immunostaining analysis, increased with the viral infection time. Morphological changes occurring during viral infection consist mainly of variations of cellular surface, different size and shape of cytoplasmic organelles and modification of nuclear components.