Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein

Matti Karp, Christer Lindqvist, Rainer Nissinen, S. Wahlbeck, Karl Åkerman, Christian Oker-Blom (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

A genetic fusion between streptavidin of Streptomyces avidinii and luciferase of Pyrophorus plagiophthalamus was constructed. The fusion protein was produced in the Sf9 insect cell line using the baculovirus expression vector system (REVS). Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the protei nsfrom cells infected with the recombinant virus, VL1393-LucGR-StreptAv, revealed that the fusion protein migrated with an apparent molecular weight of 75 kDa. Light emission measurements showed that the infected cells produced about 255 mg of the chimeric protein per liter of cell culture (127.5 μg/1 × 106 cells). Precipitation of the LucGR-StreptAv fusion protein with biotinylated acrylic beads as well as immunoblot analyses using biotinylated immunoglobulins indicated that both fusion moieties of the chimeric protein product were functional with respect to their physical and enzymatic activities.
Original languageEnglish
Pages (from-to)452-459
Number of pages8
JournalBioTechniques
Volume20
Issue number3
DOIs
Publication statusPublished - 1996
MoE publication typeA1 Journal article-refereed

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Streptavidin
Baculoviridae
Luciferases
Fusion reactions
Molecules
Proteins
Sf9 Cells
Proteus
Light emission
Streptomyces
Electrophoresis
Viruses
Cell culture
Sodium Dodecyl Sulfate
Acrylics
Insects
Immunoglobulins
Polyacrylamide Gel Electrophoresis
Cell Culture Techniques
Molecular Weight

Cite this

Karp, M., Lindqvist, C., Nissinen, R., Wahlbeck, S., Åkerman, K., & Oker-Blom, C. (1996). Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein. BioTechniques, 20(3), 452-459. https://doi.org/10.2144/19962003452
Karp, Matti ; Lindqvist, Christer ; Nissinen, Rainer ; Wahlbeck, S. ; Åkerman, Karl ; Oker-Blom, Christian. / Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein. In: BioTechniques. 1996 ; Vol. 20, No. 3. pp. 452-459.
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Karp, M, Lindqvist, C, Nissinen, R, Wahlbeck, S, Åkerman, K & Oker-Blom, C 1996, 'Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein', BioTechniques, vol. 20, no. 3, pp. 452-459. https://doi.org/10.2144/19962003452

Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein. / Karp, Matti; Lindqvist, Christer; Nissinen, Rainer; Wahlbeck, S.; Åkerman, Karl; Oker-Blom, Christian (Corresponding Author).

In: BioTechniques, Vol. 20, No. 3, 1996, p. 452-459.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Karp, Matti

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