Crossref journal-article
American Society for Microbiology
Journal of Virology (235)
Abstract

The insect baculovirus Autographa californica nuclear polyhedrosis virus was used as an expression vector for the simian virus 40 (SV40) small t (t) and large T (T) antigens. Spodoptera frugiperda (SF9) cells infected with recombinant viruses encoding these proteins produced approximately 1 to 2 micrograms of t and up to 30 micrograms of T per 3 X 10(6) cells. The former was highly soluble after Nonidet P-40 extraction of the infected cells, unlike its Escherichia coli-produced counterpart. Both SF9-produced proteins were of authentic size and could be readily immunoprecipitated by specific antibodies. Single-step immunoaffinity chromatography was used to purify the two proteins to near homogeneity, with yields averaging 70% in each case. Experiments to test the biological activity of the baculovirus SV40 proteins showed that SF9 t was capable of associating with two of the cellular proteins reported to bind to t in SV40-infected mammalian cells. Moreover, SF9 T had ATPase activity comparable to that of T produced in monkey cells, exhibited helicase activity and SV40 origin-specific DNA binding, and was active in the SV40 DNA replication assay in vitro. Thus, the SV40 T antigens produced in insect cells can be used in future studies of their biochemical roles in vitro and in vivo.

Bibliography

Murphy, C. I., Weiner, B., Bikel, I., Piwnica-Worms, H., Bradley, M. K., & Livingston, D. M. (1988). Purification and functional properties of simian virus 40 large and small T antigens overproduced in insect cells. Journal of Virology, 62(8), 2951–2959.

Authors 6
  1. C I Murphy (first)
  2. B Weiner (additional)
  3. I Bikel (additional)
  4. H Piwnica-Worms (additional)
  5. M K Bradley (additional)
  6. D M Livingston (additional)
References 0 Referenced 37

None

Dates
Type When
Created 5 years, 7 months ago (Jan. 6, 2020, 9:58 a.m.)
Deposited 3 years, 5 months ago (March 4, 2022, 6:26 p.m.)
Indexed 1 year, 10 months ago (Oct. 1, 2023, 8:49 a.m.)
Issued 37 years ago (Aug. 1, 1988)
Published 37 years ago (Aug. 1, 1988)
Published Print 37 years ago (Aug. 1, 1988)
Funders 0

None

@article{Murphy_1988, title={Purification and functional properties of simian virus 40 large and small T antigens overproduced in insect cells}, volume={62}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.62.8.2951-2959.1988}, DOI={10.1128/jvi.62.8.2951-2959.1988}, number={8}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Murphy, C I and Weiner, B and Bikel, I and Piwnica-Worms, H and Bradley, M K and Livingston, D M}, year={1988}, month=aug, pages={2951–2959} }