Highly efficient production of GFP and its derivatives in insect cells for visual in vitro applications

Christian Oker-Blom (Corresponding Author), Adelina Orellana, Kari Keinänen

Research output: Contribution to journalArticleScientificpeer-review

31 Citations (Scopus)

Abstract

We have generated recombinant baculoviruses for expression of the green fluorescent protein (GFP), a bright GFP mutant (S65T), and a GFP-streptavidin fusion protein in Sf9 and High Five insect cell lines. At 3–4 days post infection, about 30% of the total protein contents was represented by the recombinant protein products, giving the infected insect cells a bright green color which was clearly visible by eye in daylight. The isolated GFP-streptavidin fusion protein, which possessed fluorescence properties identical to those of the native GFP, was capable of binding biotin as shown by using biotinylated beads as well as biotinylated antibody complexes decorating surface expressed GluR-6 glutamate receptor in live and fixed insect cells. The exceptionally high expression levels of GFP and GFP (S65T) and the GFP-streptavidin fusion protein in recombinant baculovirus infected insects should facilitate production of GFP derivatives for in vitro applications.
Original languageEnglish
Pages (from-to)238-243
Number of pages6
JournalFEBS Letters
Volume389
Issue number3
DOIs
Publication statusPublished - 1996
MoE publication typeA1 Journal article-refereed

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Green Fluorescent Proteins
Insects
Derivatives
Streptavidin
Fusion reactions
Baculoviridae
Proteins
Recombinant Fusion Proteins
In Vitro Techniques
Glutamate Receptors
Biotin
Recombinant Proteins
Color
Fluorescence
Cells
Cell Line
Antibodies
Infection

Cite this

Oker-Blom, Christian ; Orellana, Adelina ; Keinänen, Kari. / Highly efficient production of GFP and its derivatives in insect cells for visual in vitro applications. In: FEBS Letters. 1996 ; Vol. 389, No. 3. pp. 238-243.
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Highly efficient production of GFP and its derivatives in insect cells for visual in vitro applications. / Oker-Blom, Christian (Corresponding Author); Orellana, Adelina; Keinänen, Kari.

In: FEBS Letters, Vol. 389, No. 3, 1996, p. 238-243.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Highly efficient production of GFP and its derivatives in insect cells for visual in vitro applications

AU - Oker-Blom, Christian

AU - Orellana, Adelina

AU - Keinänen, Kari

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PY - 1996

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AB - We have generated recombinant baculoviruses for expression of the green fluorescent protein (GFP), a bright GFP mutant (S65T), and a GFP-streptavidin fusion protein in Sf9 and High Five insect cell lines. At 3–4 days post infection, about 30% of the total protein contents was represented by the recombinant protein products, giving the infected insect cells a bright green color which was clearly visible by eye in daylight. The isolated GFP-streptavidin fusion protein, which possessed fluorescence properties identical to those of the native GFP, was capable of binding biotin as shown by using biotinylated beads as well as biotinylated antibody complexes decorating surface expressed GluR-6 glutamate receptor in live and fixed insect cells. The exceptionally high expression levels of GFP and GFP (S65T) and the GFP-streptavidin fusion protein in recombinant baculovirus infected insects should facilitate production of GFP derivatives for in vitro applications.

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