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Analysis of T-DNA-mediated translational β-glucuronidase gene fusions

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Abstract

Three random translational β-glucuronidase (gus) gene fusions were previously obtained in Arabidopsis thaliana, using Agrobacterium-mediated transfer of a gus coding sequence without promoter and ATG initiation site. These were analysed by IPCR amplification of the sequence upstream of gus and nucleotide sequence analysis. In one instance, the gus sequence was fused, in inverse orientation, to the nos promoter sequence of a truncated tandem T-DNA copy and translated from a spurious ATG in this sequence. In the second transgenic line, the gus gene was fused to A. thaliana DNA, 27 bp downstream an ATG. In this line, a large deletion occurred at the target site of the T-DNA. In the third line, gus is fused in frame to a plant DNA sequence after the eighth codon of an open reading frame encoding a protein of 619 amino acids. This protein has significant homology with animal and plant (receptor) serine/threonine protein kinases. The twelve subdomains essential for kinase activity are conserved. The presence of a potential signal peptide and a membrane-spanning domain suggests that it may be a receptor kinase. These data confirm that plant genes can be tagged as functional translational gene fusions.

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Author notes

  1. Sunee Kertbundit

    Present address: Molecular Genetics, Genetic Engineering Center, Institute for Science and Technology for Research and Development, Mahidol University, Salaya Campus, Phutthamonthol Sub-district, Nakorn Pathom, 73170, Thailand

  2. Rosario Linacero

    Present address: Departamento de Genetica, Facultad de Ciencias Biologicas, Universidad Complutense de Madrid, 28040, Madrid, Spain

  3. Ivan Galis & Jiri Macas

    Present address: Institute of Plant Molecular Biology, Academy of Sciences of Czech Republic (IPMB ASCR), Branisovska 31, 37005, Ceske Budejovice, Czech Republic

  4. Suzy Renckens

    Present address: Section of Biosafety and Biotechnology, Institute of Public Health, J. Wytsmanstraat 14, 1050, Brussels, Belgium

Authors and Affiliations

  1. Laboratorium Genetische Virologie, Vrije Universiteit Brussel, Paardenstraat 65, 1640, Sint-Genesius-Rode, Belgium

    Sunee Kertbundit, Rosario Linacero, Ivan Galis, Jiri Macas, Suzy Renckens, Jean-Pierre Hernalsteens & Henri De Greve

  2. Laboratoire Associé de l'INRA (France), Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), U, niversiteit Gent, Ledeganckstraat 35, 9000, Gent, Belgium

    Pierre Rouzé

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Kertbundit, S., Linacero, R., Rouzé, P. et al. Analysis of T-DNA-mediated translational β-glucuronidase gene fusions. Plant Mol Biol 36, 205–217 (1998). https://doi.org/10.1023/A:1005902730810

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