Biopolym. Cell. 2012; 28(6):449-455.
http://dx.doi.org/10.7124/bc.000135
Molecular and Cell Biotechnologies
Creation of glyphosate-resistant Brassica napus L. plants expressing DesC desaturase of cyanobacterium Synechococcus vulcanus
1Sakhno L. O., 1Gerasymenko I. M., 1Komarnitsskii I. K., 1Sheludko Y. V., 2Goldenkova-Pavlova I. V.
  1. Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
    148, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
  2. N. I. Vavilov Institute of General Genetics RAS
    3, Gubkina Str., Moscow, Russian Federation, 119991

Abstract

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 qualitative 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.
Keywords: Brassica napus, epsps, desC, licBM3, lichenase
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