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Updated: Aug 7, 2026
Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
Published on: January 30, 2019
Development of a rapid and convenient method for the quantification of HIV-1 budding
Sayuri Sakuragi1, Jun-ichi Sakuragi, Yuko Morikawa
1Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, 3-1, Yamadaoka, Suita-City, Osaka, Japan. ssakurag@biken.osaka-u.ac.jp
Abstract:
In cells, the expression of Gag protein, one of the major structural proteins of retroviruses, is sufficient for budding virus-like particles (VLPs) from the cell surface. We have previously reported that spheroplasts of Saccharomyces cerevisiae expressing HIV-1 Gag proteins from an episomal plasmid constitutively released a large amount of VLPs into culture media; however, commercially available ELISA kits which detect mature capsid of HIV-1 could not detect uncleaved 55-kDa Gag proteins released from budding yeast. We therefore developed a method to quantitate VLP levels released from budding yeast by using fusion protein from HIV-1 Gag and Firefly Luciferase. This system is useful for screening cellular factor(s) involved in retrovirus budding from S. cerevisiae.
