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

• Published in: Journal of Asia-Pacific entomology 1999-09, Vol.2 (2), p.143-148
• Main Authors: Cho, Eun Sook, Park, Hae Jin, Lee, Kwang Sik, Je, Yeon Ho, Kang, Seok Woo, Yun, Eun Young, Kim, Keun Young, Sohn, Hung Dae, Kang, Seok Kwon, Jin, Byung Rae
• Format: Article
• Language: English
• Subjects: Autographa californica; Baculovirus; Cell division; Cytoplasm; genomics; Green fluorescent protein; IE1 gene; IE1 protein; Immediate-early proteins; Insect cells; Integration; Nuclear polyhedrosis virus; Plasmids; Polymerase chain reaction; Primers; Promoters; Replication; Transformed cells
• ISSN: 1226-8615; 1876-7990
• DOI: 10.1016/S1226-8615(08)60042-4

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.