Development of a rapid and convenient method for the quantification of HIV-1 budding
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...
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| Veröffentlicht in: | Microbes and infection 2006-06, Vol.8 (7), p.1875-1881 |
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| container_title | Microbes and infection |
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| creator | Sakuragi, Sayuri Sakuragi, Jun-ichi Morikawa, Yuko Shioda, Tatsuo |
| description | 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. |
| doi_str_mv | 10.1016/j.micinf.2006.02.027 |
| format | Article |
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Psychology</subject><subject>Gene Products, gag - biosynthesis</subject><subject>Gene Products, gag - genetics</subject><subject>Gene Products, gag - metabolism</subject><subject>HIV-1</subject><subject>HIV-1 - genetics</subject><subject>HIV-1 - physiology</subject><subject>Human immunodeficiency virus 1</subject><subject>Humans</subject><subject>Luciferase</subject><subject>Luciferases, Firefly - genetics</subject><subject>Luciferases, Firefly - metabolism</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Mycology</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Spheroplasts - genetics</subject><subject>Spheroplasts - physiology</subject><subject>Virology</subject><subject>Virology - methods</subject><subject>Virosomes - metabolism</subject><issn>1286-4579</issn><issn>1769-714X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2KFDEUhYMozo--gUhtdFftTaoqPxtBRscZGHAziruQSm6cNFVJT1LV4Nubthtmp3AhIfnOIXwh5A2FDQXKP2w3c7Ah-g0D4BtgdcQzck4FV62g_c_ndc8kb_tBqDNyUcoWgA6C9y_JGeUSBHTinNx_xj1OaTdjXJrkG9NkswuuMdE1NsU9xnC4mXF5SK7xKTfLAzaPq4lL8MGaJaR4yN3c_mhpM67OhfjrFXnhzVTw9Wm9JN-vv9xf3bR3377eXn26a-3QdUsrOoo9OGfBDUxxT5mDfmTQscEIgaP3TBipuBFOAppO9HK0jitLaT0ZfHdJ3h97dzk9rlgWPYdicZpMxLQWzaWgCkD9F6RKStVJqGB_BG1OpWT0epfDbPJvTUEftOutPmrXB-0aWB1RY29P_es4o3sKnTxX4N0JMMWayWcTbShPnKxl7G_RxyOHVds-YNbF1g-w6EJGu2iXwr9f8gcxYaGU</recordid><startdate>20060601</startdate><enddate>20060601</enddate><creator>Sakuragi, Sayuri</creator><creator>Sakuragi, Jun-ichi</creator><creator>Morikawa, Yuko</creator><creator>Shioda, Tatsuo</creator><general>Elsevier SAS</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>20060601</creationdate><title>Development of a rapid and convenient method for the quantification of HIV-1 budding</title><author>Sakuragi, Sayuri ; Sakuragi, Jun-ichi ; Morikawa, Yuko ; Shioda, Tatsuo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c533t-731e40ddc0d5296f12d04b20325a77ebff27a896a7d80ea3748bcd69c117d85f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Biological and medical sciences</topic><topic>Fundamental and applied biological sciences. 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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
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Biological and medical sciences Fundamental and applied biological sciences. Psychology Gene Products, gag - biosynthesis Gene Products, gag - genetics Gene Products, gag - metabolism HIV-1 HIV-1 - genetics HIV-1 - physiology Human immunodeficiency virus 1 Humans Luciferase Luciferases, Firefly - genetics Luciferases, Firefly - metabolism Microbiology Miscellaneous Mycology Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Spheroplasts - genetics Spheroplasts - physiology Virology Virology - methods Virosomes - metabolism |
| title | Development of a rapid and convenient method for the quantification of HIV-1 budding |
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