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GrainGenes Reference Report: TAG-109-815

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Reference	TAG-109-815
Title	A large-scale study of rice plants transformed with different T-DNAs provides new insights into locus composition and T-DNA linkage configurations
Journal	Theoretical and Applied Genetics
Volume	109
Pages	815-826
Author	Afolabi AS Worland B Snape JW Vain P
Abstract	Transgenic locus composition and T-DNA linkage configuration were assessed in a population of rice plants transformed using the dual-binary vector system pGreen (T-DNA containing the bar and gus genes)/pSoup (T-DNA containing the aphIV and gfp genes). Transgene structure, expression and inheritance were analysed in 62 independently transformed plant lines and in around 4,000 progeny plants. The plant lines exhibited a wide variety of transgenic locus number and composition. The most frequent form of integration was where both T-DNAs integrated at the same locus (56% of loci). When single-type T-DNA integration occurred (44% of loci), pGreen T-DNA was preferentially integrated. In around half of the plant lines (52%), the T-DNAs integrated at two independent loci or more. In these plants, both mixed and single-type T-DNA integration often occurred concurrently at different loci during the transformation process. Non-intact T-DNAs were present in 70-78% of the plant lines causing 14-21% of the loci to contain only the mid to right border part of a T-DNA. In 53-66% of the loci, T-DNA integrated with vector backbone sequences. Comparison of transgene presence and expression in progeny plants showed that segregation of the transgene phenotype was not a reliable indicator of either transgene inheritance or T-DNA linkage, as only 60-80% of the transgenic loci were detected by the expression study. Co-expression (28% of lines) and backbone transfer (53-66% of loci) were generally a greater limitation to the production of marker-free T1 plants expressing the gene of interest than co-transformation (71% of lines) and unlinked integration (44% of loci)
Keyword	[ Hide all but 1 of 21 ] aphiv gene bar gene beta-glucuronidase gene expression gene segregation genes genetic recombination genetic transformation gfp gene green fluorescent protein gus gene inheritance (genetics) linkage (genetics) loci oryza sativa phenotype plasmid vectors rice transfer dna transgenes varieties

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