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Sindarovska Y.R.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine
TSitologiia i genetika | Year: 2010

Human interferon alpha2b gene was transiently expressed in Nicotiana excelsior plants. Fusion with N. plumbaginifolia calreticulin signal peptide for improved apoplast targeting and carrying out the expression under optimized conditions resulted in maximal interferon activity of 3.2 x 10(3) IU/g fresh weight (FW) with an average of 2.1 +/- 0.8 x 10(3) IU/g FW. It proves that N. excelsior is a suitable host for Agrobacterium-mediated transient expression of genes encoding physiologically active human proteins. The transient expression conditions optimized for GFP marker protein were confirmed to be preferable for hIFN alpha2b.


Sakhno L.O.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Gerasymenko I.M.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Komarnitsskii I.K.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Sheludko Y.V.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Goldenkova-Pavlova I.V.,Russian Academy of Sciences
Biopolymers and Cell | Year: 2012

Aim. Creation of glyphosate-resistant canola plants expressing bifunctional hybrid desC::licBM3 gene. In the hybrid gene the sequence of DesC desaturase of cyanobacterium S. vulcanus without plastid targeting was fused with the sequence of thermostable lichenase reporter LicBM3 gene. Methods. Agrobacterium tumefaciens- mediated transformation, PCR, quantitative and qualitative determination of lichenase activity, genetic analysis. Results. Transgenic canola plants, carring the enolpyruvat shikimat phosphate syntase gene (epsps), conferring on plants resistance to phosphonomethyl glycine herbicides (Roundup), as well as the desC::licBM3 gene, were selected. The presence of transgenes was confimed by multiplex PCR. The epsps gene expression in canola was shown at the transcription level, during in vitro growth and after greenhouse herbicide treatment. Activity of the licBM3 gene product as a part of hybrid protein allowed quantitative and qualiative estimation of the desaturase gene expression. Inheritance of heterologous genes and their expression in the first generation were investigated. Conclusions. Transgenic canola plants were obtained, the presence of trangenes in plant genome was proved and expression of the target genes was detected. © Institute of Molecular Biology and Genetics, NAS of Ukraine, 2012.


Sindarovska Y.R.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Golovach I.S.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Belokurova V.B.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | Gerasymenko I.M.,Institute of Cell Biology and Genetic Engineering NAS of Ukraine | And 2 more authors.
Cytology and Genetics | Year: 2014

Agrobacterium-mediated transient expression is an approach for short-time expression of heterologous genes in plant systems. During the last decade transient expression was regarded as a potent protocol for high scale production of foreign proteins in plants including pharmaceutically valuable proteins. In vitro grown plant cell cultures represent a suitable system for accumulation of heterologous proteins under controlled conditions. Since host characteristics may strongly influence transient expression efficiency, we performed screening of undifferentiated cell cultures for transient expression ability using GUS as a reporter. Analysis of 248 plant species belonging to 49 families from the National Germplasm Bank of the World Flora of the Institute of Cell Biology and Genetic Engineering (Kyiv, Ukraine) allowed for selection of about 50 plant species exhibiting detectable β-glucuronidase activity. © 2014 Allerton Press, Inc.

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