High-level expression of human interferon alpha-2b in transgenic carrot (Daucus carota L.) plants

被引:35
|
作者
Luchakivskaya, Yu. [1 ]
Kishchenko, O. [1 ]
Gerasymenko, I. [1 ]
Olevinskaya, Z. [2 ]
Simonenko, Yu. [1 ]
Spivak, M. [2 ]
Kuchuk, M. [1 ]
机构
[1] Natl Acad Sci Ukraine, Inst Cell Biol & Genet Engn, UA-03680 Kiev, Ukraine
[2] Natl Acad Sci Ukraine, Danilo Zabolotny Inst Microbiol & Virol, UA-03680 Kiev, Ukraine
关键词
Human interferon alpha-2b; Carrot plants; Agrobacterium-mediated transformation; Antiviral activity; Recombinant proteins; HUMAN GROWTH-HORMONE; NEUTRALIZING IMMUNOGENICITY; ENHANCED RESISTANCE; SUSPENSION CELLS; GENE-EXPRESSION; TRANSFORMATION; PROTEIN; ALPHA; CHITINASE; LOCALIZATION;
D O I
10.1007/s00299-010-0942-5
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
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 x 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 x 10(3) IU/g FW) and could possibly be used for generating interferon alpha-2b protein in planta for preventing and curing infectious diseases.
引用
收藏
页码:407 / 415
页数:9
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