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TAXI : a new vehicle for the transfer of genes into monocotyledonous plants

Bibliography: leaves 90-103.

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Main Author: Chen, Wusi
Other Authors: Von Holt, Claus
Format: Thesis
Language:English
Published: Department of Molecular and Cell Biology 2014
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access_status_str Open Access
author Chen, Wusi
author2 Von Holt, Claus
author_browse Chen, Wusi
Von Holt, Claus
author_facet Von Holt, Claus
Chen, Wusi
author_sort Chen, Wusi
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description Bibliography: leaves 90-103.
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institution University of Cape Town (South Africa)
language eng
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2014
publishDateRange 2014
publishDateSort 2014
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spelling oai:open.uct.ac.za:11427/9242 TAXI : a new vehicle for the transfer of genes into monocotyledonous plants Chen, Wusi Von Holt, Claus Thomson, Jennifer Ann Klump, HH Biochemistry Bibliography: leaves 90-103. The transfer of foreign genes into cereals followed by their correct expression in a tissue specific, developmentally regulated manner has become an important research focus. Based on the mechanism of Agrobacterium tumefaciens mediated transformation in dicots, a modified method to transform the monocotyledonous rye ( Secale cereale L. ) via A.tumefaciens mediated transformation was attempted. The induced bacterium culture was injected into rye seedlings, the transferred reporter gene, uid.A, was detected by PCR, and the expression of the gene was tested by histochemical assays. However, successful transformation and integration of the transgenes remained doubtful, because the frequency of kanamycin resistance in the progenies ( Rl , R2 and R3 ) did not increase. To achieve a real transformation and heritable transgenic rye, a new vehicle for gene transfer to plants was developed. A macromolecular complex, termed the TAXI, consisted of histone HI-protected single stranded DNA, containing a selectable marker gene (npt JI), linked either to a reporter gene (uidA) or a glutenin gene. The constructs were transferred by injection of rye seedlings. Molecular analyses demonstrated that all three genes were integrated and expressed in transformed rye and their progenies ( Rl and R2 ). TAXI mediated gene transfer to rye revealed an important advantage in that single or low numbers of transgenes were inserted into the transformed plant genome. However, the method of TAXI delivery to plants was not efficient. To improve this, a new approach, combining TAXI transformation and the biolistic process, was developed. A rapid regenerable callus line of a grass species, Digitaria sanguinalis, was established as a test system. TAXI coated gold particles, carrying a selectable marker gene (bar) and a reporter gene (uid.A), were used in bombardment experiments. The results of herbicide resistance and molecular analyses demonstrated that single copies or low numbers of the bar gene were inserted and expressed in regenerated transformed D. sanguinalis. Mendelian segregation in the Rl population was observed in four out of five transgenic lines. 2014-11-05T17:31:22Z 2014-11-05T17:31:22Z 1996 Doctoral Thesis Doctoral PhD http://hdl.handle.net/11427/9242 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town
spellingShingle Biochemistry
Chen, Wusi
TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
thesis_degree_str Doctoral
title TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
title_full TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
title_fullStr TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
title_full_unstemmed TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
title_short TAXI : a new vehicle for the transfer of genes into monocotyledonous plants
title_sort taxi a new vehicle for the transfer of genes into monocotyledonous plants
topic Biochemistry
url http://hdl.handle.net/11427/9242
work_keys_str_mv AT chenwusi taxianewvehicleforthetransferofgenesintomonocotyledonousplants