Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors |
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Authors: | Juan M Vega Weichang Yu Fangpu Han Akio Kato Eric M Peters Zhanyuan J Zhang James A Birchler |
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Institution: | (1) Division of Biological Sciences, University of Missouri, Tucker Hall, Columbia, MO 65211, USA;(2) Present address: Departamento de Genética, Facultad de Biología, Universidad Complutense, 28040 Madrid, Spain;(3) Present address: Faculty of Agriculture, Kyoto Prefectural University, Sakyo-ku, Kyoto-shi Kyoto-fu, 606-0823, Japan;(4) Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, Columbia, MO 65211, USA |
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Abstract: | The Cre/loxP site-specific recombination system has been applied in various plant species including maize (Zea mays) for marker gene removal, gene targeting, and functional genomics. A BIBAC vector system was adapted for maize transformation
with a large fragment of genetic material including a herbicide resistance marker gene, a 30 kb yeast genomic fragment as
a marker for fluorescence in situ hybridization (FISH), and a 35S-lox-cre recombination cassette. Seventy-five transgenic lines were generated from Agrobacterium-mediated transformation of a maize Hi II line with multiple B chromosomes. Eighty-four inserts have been localized among
all 10 A chromosome pairs by FISH using the yeast DNA probe together with a karyotyping cocktail. No inserts were found on
the B chromosomes; thus a bias against the B chromosomes by the Agrobacterium-mediated transformation was revealed. The expression of a cre gene was confirmed in 68 of the 75 transgenic lines by a reporter construct for cre/lox mediated recombination. The placement of the cre/lox site-specific recombination system in many locations in the maize genome will be valuable materials for gene targeting and
chromosome engineering. |
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Keywords: | Agrobacterium-mediated transformation B chromosome BIBAC vector Cre/loxP Maize |
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