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1999•Journal of Veterinary Medical ScienceOpen access

Expression of Bovine Interleukin-2 in Insect Cells by Recombinant Baculovirus

Takayuki Kashima, Hiroyuki Iwata, Takeshi Inoue

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Abstract

Bovine interleukin-2 (bIL-2) was expressed in insect cells infected with recombinant baculovirus. The cDNA for bIL-2 was amplified by reverse transcriptional polymerase chain reaction and recombinant baculovirus was constructed by homologous recombination. The recombinant baculovirus was plaque-purified, amplified, and then infected to Sf9 cells for expression of recombinant bIL-2 (rbIL-2). In result, the protein band corresponding to rbIL-2 could be detected on SDS-PAGE with coomassie brilliant blue staining and on immunoblot analysis reacted with mouse antiserum against bIL-2. In addition, cell proliferation assay for bIL-2 activity demonstrated that the culture supernatant of Sf9 cells infected with recombinant baculovirus enhanced the proliferation of bovine peripheral blood mononuclear cells in a dose-dependent manner.

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Bovine interleukin-2 (bIL-2) was expressed in insect cells infected with recombinant baculovirus. The cDNA for bIL-2 was amplified by reverse transcriptional polymerase chain reaction and recombinant baculovirus was constructed by homologous recombination. The recombinant baculovirus was plaque-purified, amplified, and then infected to Sf9 cells for expression of recombinant bIL-2 (rbIL-2). In result, the protein band corresponding to rbIL-2 could be detected on SDS-PAGE with coomassie brilliant blue staining and on immunoblot analysis reacted with mouse antiserum against bIL-2. In addition, cell proliferation assay for bIL-2 activity demonstrated that the culture supernatant of Sf9 cells infected with recombinant baculovirus enhanced the proliferation of bovine peripheral blood mononuclear cells in a dose-dependent manner.

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Available abstract

Bovine interleukin-2 (bIL-2) was expressed in insect cells infected with recombinant baculovirus. The cDNA for bIL-2 was amplified by reverse transcriptional polymerase chain reaction and recombinant baculovirus was constructed by homologous recombination. The recombinant baculovirus was plaque-purified, amplified, and then infected to Sf9 cells for expression of recombinant bIL-2 (rbIL-2). In result, the protein band corresponding to rbIL-2 could be detected on SDS-PAGE with coomassie brilliant blue staining and on immunoblot analysis reacted with mouse antiserum against bIL-2. In addition, cell proliferation assay for bIL-2 activity demonstrated that the culture supernatant of Sf9 cells infected with recombinant baculovirus enhanced the proliferation of bovine peripheral blood mononuclear cells in a dose-dependent manner.

Key concepts: Sf9, Recombinant DNA, Molecular biology, Baculoviridae, Complementary DNA, Biology, Antiserum, Coomassie Brilliant Blue

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Expression of Bovine Interleukin-2 in Insect Cells by Recombinant Baculovirus — Research Paper | ScholarLens