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

• Main Authors: Sunee Kertbundit, Rosario Linacero, Pierre Rouzé, Ivan Galis, Jiri Macas, Francine Deboeck, Suzy Renckens, Jean Pierre Hernalsteens, Henri De Greve
• Other Authors: Vrije Universiteit Brussel
• Format: Article
• Published: 2018
• Subjects: Biochemistry, Genetics and Molecular Biology
• Online Access: https://repository.li.mahidol.ac.th/handle/123456789/18331
• Institution: Mahidol University

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 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 suggest that it may be a receptor kinase. These data confirm that plant genes can be tagged as functional translational gene fusions.