Marker gene elimination from transgenic barley, using co-transformation with adjacent 'twin T-DNAs' on a standard Agrobacterium transformation vector
Data(s) |
2001
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Resumo |
We have tested a methodology for the elimination of the selectable marker gene after Agrobacterium-mediated transformation of barley. This involves segregation of the selectable marker gene away from the gene of interest following co-transformation using a plasmid carrying two T-DNAs, which were located adjacent to each other with no intervening region. A standard binary transformation vector was modified by insertion of a small section composed of an additional left and right T-DNA border, so that the selectable marker gene and the site for insertion of the gene of interest (GOI) were each flanked by a left and right border. Using this vector three different GOIs were transformed into barley. Analysis of transgene inheritance was facilitated by a novel and rapid assay utilizing PCR amplification from macerated leaf tissue. Co-insertion was observed in two thirds of transformants, and among these approximately one quarter had transgene inserts which segregated in the next generation to yield selectable marker-free transgenic plants. Insertion of non-T-DNA plasmid sequences was observed in only one of fourteen SMF lines tested. This technique thus provides a workable system for generating transgenic barley free from selectable marker genes, thereby obviating public concerns regarding proliferation of these genes. |
Identificador | |
Publicador |
Springer Netherlands |
Relação |
DOI:10.1023/A:1011333321893 Matthews, Peter R., Wang, Ming-Bo, Waterhouse, Peter M., Thornton, Sarah, Fieg, Sarah J., Gubler, Frank, & Jacobsen, John V. (2001) Marker gene elimination from transgenic barley, using co-transformation with adjacent 'twin T-DNAs' on a standard Agrobacterium transformation vector. Molecular Breeding, 7(3), pp. 195-202. |
Direitos |
Copyright 2001 Springer Netherlands |
Fonte |
School of Earth, Environmental & Biological Sciences; Science & Engineering Faculty |
Palavras-Chave | #060400 GENETICS #060700 PLANT BIOLOGY #Agrobacterium transformation #Barley #Co-transformation #Marker-free #Selectable marker #Transgene segregation #binary vector #DNA transfer #gene insertion #gene segregation #genetic line #genetic transformation #inheritance #marker gene #plant biotechnology #plant breeding #plant leaf #polymerase chain reaction #transgenic plant #Agrobacterium #Hordeum vulgare subsp. vulgare #Rhizobium |
Tipo |
Journal Article |