Construction of Stably Transformed Bm5 and Sf9 Cells Displaying Green Fluorescence by Using Autographa californica Nuclear Polyhedrosis Virus IE1 Gene

Abstract

Transformed Bm5 or Sf9 cells displaying green fluorescence were constructed by using Autographa californica nuclear polyhedrosis virus (AcNPV) immediate early gene ( ie 1). Green fluorescent protein ( gfp) gene was introduced under the control of the AcNPV ie 1 promoter to yield expression plasmid pAcIE1-GFP. It was transfected into Sf9 or Bm5 cells and cell clones expressing GFP were selected by fluorescence microscopy. Genomic DNA from transformed cells was isolated and integration of AcNPV ie 1 gene harboring gfp gene was confirmed by PCR using AcNPV ie 1 gene-specific primers. The GFP was successfully expressed in the cytoplasm of insect cells transformed with pAcIEI-GFP and the expressed GFP was maintained during cell division. Furthermore, GFP expression by AcNPV ie 1 promoter in transformed cells was not interfered with viral replication. This suggests that transformed cells displaying foreign gene product by using AcNPV ie 1 promoter will be useful for the diverse applications of the insect cells.