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Volume 76, Issue 1

Development of a bovine adenovirus type 3-based expression vector Free

Abstract

SUMMARY

We constructed a non-defective bovine adenovirus type 3 recombinant (BAd3-Luc) containing the firefly luciferase gene inserted in the early region 3 (E3) of the BAd3 genome. Deletion of a 696 bp I–I E3 segment and insertion of the luciferase gene in E3 was confirmed by Southern blot analyses. Luciferase was expressed in Madin-Darby bovine kidney cells infected with BAd3-Luc as measured by enzymic assays and Western blotting. Analyses of luciferase expression in the presence or absence of 1---arabinofuranosyl-cytosine indicated that approximately 70–75% of luciferase expression was dependent on viral DNA replication, suggesting that transcription of the gene was at least partially under the control of a late promoter. Although yields of infectious virus for BAd3-Luc were approximately 10-fold lower than for wild-type virus, replication of the vector was still relatively efficient. In a Western blot experiment, anti-luciferase antibody reacted with a 62 kDa protein which is of the same molecular mass as the purified firefly luciferase polypeptide. Luciferase was also expressed in the 293 cell line infected with BAd3-Luc for at least 6 days postinfection as monitored by luciferase assays. Based on these observations we suggest that BAd-based expression vectors should have excellent potential for the development of recombinant vaccines for cattle and may also be suitable as vectors for gene transfer into human cells.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology 1995
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1995-01-01
2024-04-20
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Development of a bovine adenovirus type 3-based expression vector
J. Gen. Virol. 76, 93 (1995); https://doi.org/10.1099/0022-1317-76-1-93
/content/journal/jgv/10.1099/0022-1317-76-1-93
/content/journal/jgv/10.1099/0022-1317-76-1-93
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