Construction of Artemisia vulgaris L. hair root culture with human interferon alpha 2b gene / Drobot K.O. et al. 2015

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Drobot K.O., Shakhovsky A.M., Matvieieva N.A.

PCR analysis of genes nptII (1-3), rolB (4-6) and ifn-α2b (7– 9) in transgenic roots of mugwort, otained by means of A. rhizogenes with рСВ161 vector: 1, 4, 7 – DNA of nontransgenic plants; 2, 3, 5, 6, 8, 9 – DNA of transgenic roots; M – markers

Aims. The obtaining of mugwort Artemisia vulgaris L. “hairy” root culture with human interferon α2b gene (ifn-α2b) was the aim of this work. Methods. We used Agrobacterium rhizogenes wild strain A4 as well as A. rhizogenes carried pCB161 vector with ifn-α2b gene under constitutive Mll rootspecific sugar beet promoter for mugwort genetic transformation. The genes were transferred into leaf and root explants. The presence of transgenes were determined by PCR analysis. Results. Transformation frequency was up to 100 % in case of leaf explants. Transgenic roots from other type of explants were not obtained. PCR analysis proved the presence of nptII, ifn-α2b and rolB genes in mugwort “hairy” roots obtained via A. rhizogenes-mediated transformation. Conclusions. Thus, we obtained the transgenic Artemisia vulgaris L. “hairy” roots using A. rhizogenes-mediated transformation and transferred ifn-α2b. Transformational frequency was the highest in case of leaves using as an explants – up to 100 %.

Keywords: genetic transformation, Agrobacterium rhizogenes, hairy roots, Artemisia vulgaris L., ifn-α2b gene.

PCR analysis of genes rolB (4-6) in transgenic roots of mugwort, tranformated with wild strain of A. rhizogenes A4; 1–9 – DNA of “hairy” roots, 10 – DNA of nontransgenic plants; 11 – plasmid DNA; М – marker
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