Query (optional)   in Class  All Allele Author Colleague Collection Assembly Gene Gene Class Gene Product Germplasm Image Journal Keyword Library Locus Map Map Data Marker Pathology Polymorphism Probe Protein QTL Rearrangement Reference Sequence Species Trait Trait Study Two Point Data  

---

GrainGenes Reference Report: MGG-239-354

[Submit comment/correction]

Reference	MGG-239-354
Title	Trans-activation and stable integration of the maize transposable element Ds cotransfected with the Ac transposase gene in transgenic rice plants
Journal	Molecular and General Genetics
Year	1993
Volume	239
Pages	354-360
Author	Shimamoto K [ Show all 8 ]
Abstract	To develop an efficient gene tagging system in rice, a plasmid was constructed carrying a non-autonomous maize Ds element in the untranslated leader sequence of a hygromycin B resistance gene fused with the 35S promoter of cauliflower mosaic virus This plasmid was cotransfected by electroporation into rice protoplasts together with a plasmid containing the maize Ac transposase gene transcribed from the 35S promoter Five lines of evidence obtained from the analyses of hygromycin B-resistant calli, regenerated plants and their progeny showed that the introduced Ds was trans-activated by the Ac transposase gene in rice (1) Cotransfection of the two plasmids is necessary for generation of hygromycin B resistant transformants (2) Ds excision sites are detected by Southern blot hybridization (3) Characteristic sequence alterations are found at Ds excision sites (4) Newly integrated Ds is detected in the rice genome. (5) Generation of 8 bp target duplications is observed at the Ds Integration sites on the rice chromosomes. Our results also show that Ds can be trans-activated by the transiently expressed Ac transposase at early stages of protoplast culture and integrated stably into the rice genome, while the cotransfected Ac transposase gene is not integrated. Segregation data from such a transgenic rice plant carrying no Ac transposase gene showed that four Ds copies were stably integrated into three different chromosomes, one of which also contained the functional hph gene restored by Ds excision. The results indicate that a dispersed distribution of Ds throughout genomes not bearing the active Ac transposase gene can be achieved by simultaneous transfection with Ds and the Ac transposase gene.
Keyword	[ Hide all but 1 of 21 ] direct-dna-uptake drug-resistance duplication electroporation gene-interaction gene-tagging genetic change genetic transformation hph gene hygromycin-b hygromycin-b-phosphotransferase insertional-mutagenesis oryza sativa phosphotransferases plasmid-vectors reporter genes segregation transgenic-plants transposable-elements transposition zea mays

GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.