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Expression of recombinant Digitalis lanata EHRH. cardenolide 16′-O-glucohydrolase in Cucumis sativus L. hairy roots

  • Genetic Transformation and Hybridization
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Abstract

The coding sequence for the Digitalis lanata EHRH. cardenolide 16′-O-glucohydrolase was inserted downstream of the 35S promoter in the binary vector pBI121 resulting in plant expression vector pBI121cgh. Cotyledon explants excised from 10-day-old seedlings of Cucumis sativus L. were transformed using Agrobacterium rhizogenes 15834 harbouring pBI121cgh. Hairy roots were obtained from infected cotyledon explants in vitro 10 days after inoculation. PCR amplification of coding sequences for cgh I, rolB and rolC from Ri plasmid showed that the aimed sequences were inserted into the genome of transformed cucumber hairy roots. Glycolytic activity of the transgenic CGH I was measured by HPLC using Lanatoside glycosides as substrate. Therefore, the cgh I transformed cucumber hairy roots may provide a valuable model for biotransformation of natural compounds by recombinant enzymes.

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Abbreviations

CGHI:

Cardenolide 16′-O-glucohydrolase

PCR:

Polymerase chain reaction

MS:

Murashige and Skoog

HPLC:

High-Performance Liquid chromatography

Ri:

Root inducing

rolB :

Rooting locus B

rolC :

Rooting locus C

YEB:

Yeast extract broth

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Acknowledgement

A DAAD fellowship for Shi He-Ping is acknowledged.

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Author notes

  1. He-Ping Shi

    Present address: College of Life Science, South China Normal University, Guangzhou, 510631, P. R. China

Authors and Affiliations

  1. Institut für Pharmakologie und Pharmazeutische Biologie, Martin-Luther—Universität, Hoher Weg 8, 06120, Halle (Saale), Germany

    He-Ping Shi & Peter Lindemann

Authors
  1. He-Ping Shi
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  2. Peter Lindemann
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Corresponding author

Correspondence to Peter Lindemann.

Additional information

Communicated by H. Lörz

This report is dedicated to Prof. W. Roos on the occasion of his 60th birthday.

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Shi, HP., Lindemann, P. Expression of recombinant Digitalis lanata EHRH. cardenolide 16′-O-glucohydrolase in Cucumis sativus L. hairy roots. Plant Cell Rep 25, 1193–1198 (2006). https://doi.org/10.1007/s00299-006-0183-9

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  • DOI: https://doi.org/10.1007/s00299-006-0183-9

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