A functional hepatitis B virus X protein produced in insect cells

Dan Spandau, Hwei gene Heidi Wang, Malcolm J. Fraser, Chao-Hung Lee

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We have constructed a recombinant baculovirus from Autographa californica nuclear polyhedrosis virus, called AcX, that expresses the gene encoding the hepatitis B virus X protein in infected Spodoptera frugiperda (Sf21AE) insect cells. A 16.5-kDa monomer and a 33-kDa dimer of the X protein were detected in extracts from AcX-infected cells on immunoblots using a polyclonal anti-X antibody. The biological activity of the insect cell-produced X protein was assayed by fusing AcX-infected Sf21 AE cells with African green monkey kidney (CV-1) cells and then transfecting the fused cells with the reporter plasmid pSV2cat. The expression of the cat gene in CV- 1:Sf21AE(AcX) fusions was seven times higher than that derived from CV-1:Sf21AE(AcMNPV) fusions, indicating that the insect cell-produced X protein was functional. The transactivation function of the insect cell-produced X protein was also verified by scrape-loading nuclear extracts of AcX-infected Sf21AE cells into pSV2cat-transfected CV-1 cells. Treatment of the AcX-infected cell nuclear extracts with anti-X antisera prior to scrape-loading eliminated the transactivating activity of the extracts. We conclude that the insect cell-produced X protein was functionally identical to that generated in mammalian cells.

Original languageEnglish
Pages (from-to)938-941
Number of pages4
JournalVirology
Volume185
Issue number2
DOIs
StatePublished - 1991

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Insects
Proteins
hepatitis B virus X protein
Nucleopolyhedrovirus
Sf9 Cells
Cercopithecus aethiops
Spodoptera
Baculoviridae
Cell Extracts
Transcriptional Activation
Immune Sera
Anti-Idiotypic Antibodies
Cats
Plasmids
Kidney
Gene Expression
1,2-diamino-1,2-N,N'-carbonyl-1,2-dideoxyglucose hydrate

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

A functional hepatitis B virus X protein produced in insect cells. / Spandau, Dan; Wang, Hwei gene Heidi; Fraser, Malcolm J.; Lee, Chao-Hung.

In: Virology, Vol. 185, No. 2, 1991, p. 938-941.

Research output: Contribution to journalArticle

Spandau, Dan ; Wang, Hwei gene Heidi ; Fraser, Malcolm J. ; Lee, Chao-Hung. / A functional hepatitis B virus X protein produced in insect cells. In: Virology. 1991 ; Vol. 185, No. 2. pp. 938-941.
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