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: GNM-44-691

[Submit comment/correction]

Reference	GNM-44-691
Title	Genomic interspersions determine the size and complexity of transgene loci in transgenic plants produced by microprojectile bombardment
Journal	Genome
Year	2001
Volume	44
Pages	691-697
Author	Svitashev SK Somers DA
Abstract	Summary: The structure of transgene loci in six transgenic allohexaploid oat (Avena sativa L.) lines produced using microprojectile bombardment was characterized using fluorescence in situ hybridization (FISH) on extended DNA fibers (fiber-FISH). The transgene loci in five lines were composed of multiple copies of delivered DNA interspersed with genomic DNA fragments ranging in size from ca. 3 kb to at least several hundred kilobases, and in greater numbers than detected using Southern blot analysis. Although Southern analysis predicted that the transgene locus in one line consisted of long tandem repeats of the delivered DNA, fiber-FISH revealed that the locus actually contained multiple genomic interspersions. These observations indicated that transgene locus size and structure were determined by the number of transgene copies and, possibly to a greater extent, the number and the length of interspersing genomic DNA sequences within the locus. Large genomic interspersions detected in several lines were most likely the products of chromosomal breakage induced either by tissue culture conditions or, more likely, by DNA delivery into the nucleus using microprojectile bombardment. We propose that copies of transgene along with other extrachromosomal DNA fragments are used as patches to repair double-strand breaks (DSBs) in the plant genome resulting in the formation of transgene loci
Keyword	[ Hide all but 1 of 53 ] arabidopsis thaliana avena avena sativa bombardment breakage canada chromosome complexity culture culture conditions delivery dna dna delivery dna fragments dna sequence double-strand break extended dna fibers fiber fish fluorescence fluorescence in situ hybridization fragments genetic genetic transformation genome genomic dna hybridization in situ in situ hybridization inheritance line loci locus microprojectile bombardment nucleus oat patch plant genome repair repeat sequence situ hybridization structure tandem tandem repeat tissue tissue culture transformation transgene transgene locus structure transgenic transgenic oat transgenic plant

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