High-level expression of human interferon alpha-2b in transgenic carrot (Daucus carota L.) plants.
In this study, we report the obtaining of carrot plants expressing human interferon alpha-2b via Agrobacterium-mediated transformation using two vector constructs containing the sequence coding for interferon gene fused with Nicotiana plumbagenifolia calreticulin apoplast targeting signal driven by 35S CaMV promoter and root-specific Mll promoter. The human interferon alpha-2b gene was correctly translated in carrot plants according to Western blot analysis. The recombinant protein exhibited antiviral activity in vitro by inhibition of vesicular stomatitis virus replication in established piglet testicular cells. The results demonstrated the higher activity of interferon accumulated in carrot plants for young leaves (up to 50.7 × 10(3) IU/g FW) compared to the mature ones probably due to the degradation-susceptible nature of this protein. The taproot-expressing system could have also provided the sufficient protein amounts (up to 16.5 × 10(3) IU/g FW) and could possibly be used for generating interferon alpha-2b protein in planta for preventing and curing infectious diseases.
Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Zabolotnogo str 148, Kyiv 03680, Ukraine. yu.luchakivskaya@gmail.com
Yu
Y
Luchakivskaya
O
O
Kishchenko
I
I
Gerasymenko
Z
Z
Olevinskaya
Yu
Y
Simonenko
M
M
Spivak
M
M
Kuchuk
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D018744:DNA, Plant / Q000235:genetics; D018552:Daucus carota / Q000235:genetics*; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000494:pharmacology; D018515:Plant Leaves / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D012231:Rhizobium; D013552:Swine; D014170:Transformation, Genetic
D000998:Antiviral Agents; D018744:DNA, Plant; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins