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Gene insertion into Hevea brasiliensis

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Abstract

A transformation system has been developed for Hevea brasiliensis using the particle gun method. Anther derived calluses were transformed with vectors harbouring the ß-glucuronidase (gus) gene, the neomycin phosphotransferase (nptII) gene, and the chloramphenicol acetyl transferase (cat) gene. Gene transfer was determined by histochemical staining and fluorometric assay for ß-glucuronidase activity, enzyme linked immunosorbent assay for detecting neomycin phosphotransferase II gene and direct enzyme assay for detection of expression of the cat gene. These independent assays all showed a several-fold increase, compared to control values, in gene product level and enzyme activity in extracts from transformed callus and embryoids of Hevea. These results were confirmed using polymerase chain reaction with primers designed to amplify an internal gus fragment. Together, the results show the feasibility of the particle gun method for the introduction of foreign genes into Hevea.

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Abbreviations

BSA:

bovine serine albumin

CAT:

chloramphenicol acetyl transferase

ELISA:

enzyme-linked immunosorbent assay

GUS:

ß-glucuronidase

kb:

kilobase

MU:

4-methyl-umbelliferyl-ß-D-glucuronide

NPT-II:

neomycin phosphotransferase II

PCR:

polymerase chain reaction

Tris:

Trizma base

X-gluc:

5-bromo-4-chloro-3-indolyl glucuronide

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Authors and Affiliations

  1. Division of Life Sciences, Kings College London, Campden Hill Road, W8 7AH, London, UK

    P. Arokiaraj, S. Coomber & B. V. Charlwood

  2. Division of Biosciences, University of Hertfordshire, Hatfield Campus, College Lane, AL10 9AB, Hatfield, Herts, UK

    H. Jones

  3. Biotechnology, Biochemistry, Molecular and Cell Biology Division, Rubber Research Institute of Malaysia, Rubber Research Institute, Experimental Station, 47000, Sungei Buloh, Selangor Darul Ehsan, Malaysia

    P. Arokiaraj & K. F. Cheong

Authors
  1. P. Arokiaraj
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  2. H. Jones
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  3. K. F. Cheong
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  4. S. Coomber
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  5. B. V. Charlwood
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Additional information

Communicated by M. R. Davey

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Arokiaraj, P., Jones, H., Cheong, K.F. et al. Gene insertion into Hevea brasiliensis . Plant Cell Reports 13, 425–431 (1994). https://doi.org/10.1007/BF00231960

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  • DOI: https://doi.org/10.1007/BF00231960

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