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PMID: 1692347 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Human immunodeficiency virus type 1 Pr55gag and Pr160gag-pol expressed from a simian virus 40 late replacement vector are efficiently processed and assembled into viruslike particles.

Journal of virology ·Vol. 64 ·No. 6 ·1990-06-00 ·Pages 2743-50

Smith AJ, Cho MI, Hammarskjöld ML, Rekosh D

Abstract

Human immunodeficiency virus type 1 (HIV-1) gag and pol genes were expressed by using fragments of the BH10 clone of HIV inserted into a simian virus 40 late replacement vector. An initial construct containing the entire coding regions of gag, pol, and vif produced only minute amounts of the gag precursor, Pr55gag. However, high-level expression was obtained when an additional sequence from the env gene (the rev-responsive element) was inserted 3' of vif in the correct orientation, and rev was provided in trans from a second vector. Western immunoblot analysis of transfected cells showed the presence of large amounts of both Pr55gag and Pr160gag-pol as well as all of the expected cleavage products. Electron microscopy of thin sections of transfected cells showed a multitude of viruslike particles. Both immature particles in the process of budding and particles containing the condensed core characteristic of HIV were observed. Analysis of the released viruslike particles showed the presence of active reverse transcriptase. Sucrose gradient analysis of particles produced from [3H]uridine-labeled cells indicated a peak of radioactivity which cosedimented with a peak of p24, suggesting that the particles contained RNA.

MeSH Terms
Animals Blotting, Western Cell Line Fusion Proteins, gag-pol/genetics Gene Products, gag/genetics Genes, gag Genes, pol Genetic Vectors HIV-1/genetics,ultrastructure Microscopy, Electron Plasmids RNA-Directed DNA Polymerase/metabolism Simian virus 40/genetics Transfection
Chemicals
Fusion Proteins, gag-pol Gene Products, gag RNA-Directed DNA Polymerase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Smith A J
Department of Microbiology, State University of New York, Buffalo 14214.
Cho M I
Hammarskjöld M L
Rekosh D
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.6.2743-2750.1990
Published
1990-06-00
Pages
2743-50
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249454
Subset
IM
Grants
NIAID NIH HHS · AI25721 · United States
NCI NIH HHS · CA00905 · United States
External Links
PubMed source DOI: 10.1128/JVI.64.6.2743-2750.1990 PMC Full Text View journal details
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