Efficient expression of histidine-tagged large hepatitis delta antigen in baculovirus-transduced baby hamster kidney cells.
OBJECTIVE
To study the baculovirus/mammalian cell system for efficient expression of functional large hepatitis delta antigen (L-HDAg).


METHODS
A recombinant baculovirus expressing histidine-tagged L-HDAg (L-HDAgH) was constructed to transduce baby hamster kidney (BHK) cells by a simplified transduction protocol.


RESULTS
The recombinant baculovirus transduced BHK cells with efficiencies higher than 90% as determined by flow cytometry. The expression level was significantly higher than that obtained by plasmid transfection and was further enhanced 3-fold to around 19 pg/cell by the addition of 10 mmol/L sodium butyrate. Importantly, the expressed L-HDAgH was localized to the cell nucleus and correctly isoprenylated as determined by immunofluorescence labeling and confocal microscopy. Moreover, L-HDAgH interacted with hepatitis B surface antigen to form virus-like particles.


CONCLUSIONS
The fusion with histidine tags as well as overexpression of L-HDAgH in the baculovirus-transduced BHK cells does not impair the biological functions. Taken together, the baculovirus/mammalian cell system offers an attractive alternative for high level expression of L-HDAgH or other proteins that require extensive post-translational modifications.
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan, China.
Ying-Wei
YW
Chiang
Jaw-Chin
JC
Wu
Kuei-Chun
KC
Wang
Chia-Wei
CW
Lai
Yao-Chi
YC
Chung
Yu-Chen
YC
Hu
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D006224:Cricetinae; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000032:analysis; D006518:Hepatitis Viruses / Q000235:genetics / Q000276:immunology; D035923:Hepatitis delta Antigens / Q000235:genetics*; D006639:Histidine; D007668:Kidney; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins; D014161:Transduction, Genetic
D004274:DNA, Recombinant; D004279:DNA, Viral; D035923:Hepatitis delta Antigens; D011994:Recombinant Proteins; C442244:hepatitis delta virus large antigen; D006639:Histidine