PMID- 11135106
OWN - NLM
STAT- MEDLINE
DCOM- 20010222
LR  - 20190906
IS  - 0960-7412 (Print)
IS  - 0960-7412 (Linking)
VI  - 24
IP  - 6
DP  - 2000 Dec
TI  - Widely separated multiple transgene integration sites in wheat chromosomes are 
      brought together at interphase.
PG  - 713-23
AB  - We have investigated the organization of transgenes delivered by particle 
      bombardment into the wheat genome, combining conventional molecular analysis with 
      fluorescence in situ hybridization (FISH) and three-dimensional confocal 
      microscopy. We selected a representative population of transformed wheat lines 
      and carried out molecular and expression analysis. FISH on metaphase chromosomes 
      showed that transgene integration sites were often separated by considerable 
      lengths of genomic DNA (>1 Mbp), or could even be on opposite chromosome arms. 
      Plants showing multiple integration sites on a single chromosome were selected 
      for three-dimensional confocal analysis of interphase nuclei in root and embryo 
      tissue sections. Confocal microscopy revealed that these sites lay in close 
      physical proximity in the interphase nuclei. Our results clearly show that 
      multiple transgenes physically separated by large intervening regions of 
      endogenous DNA at metaphase can be brought together at interphase. This may 
      reflect the original physical organization of the endogenous DNA at the moment of 
      transformation, with DNA strand breaks introduced into several co-localized DNA 
      loops by the intruding gold particles. Alternatively, the transgenes may be 
      brought together after transformation, either by an ectopic homologous pairing 
      mechanism, or by recruitment to a common transcription site.
FAU - Abranches, R
AU  - Abranches R
AD  - John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.
FAU - Santos, A P
AU  - Santos AP
FAU - Wegel, E
AU  - Wegel E
FAU - Williams, S
AU  - Williams S
FAU - Castilho, A
AU  - Castilho A
FAU - Christou, P
AU  - Christou P
FAU - Shaw, P
AU  - Shaw P
FAU - Stoger, E
AU  - Stoger E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Plant J
JT  - The Plant journal : for cell and molecular biology
JID - 9207397
SB  - IM
MH  - Chromosome Mapping
MH  - Gene Transfer Techniques
MH  - In Situ Hybridization, Fluorescence
MH  - *Interphase
MH  - Microscopy, Confocal
MH  - Plant Roots
MH  - Plants, Genetically Modified
MH  - *Recombination, Genetic
MH  - Seeds
MH  - Transformation, Genetic
MH  - *Transgenes
MH  - Triticum/*genetics
EDAT- 2001/01/03 11:00
MHDA- 2001/03/03 10:01
CRDT- 2001/01/03 11:00
PHST- 2001/01/03 11:00 [pubmed]
PHST- 2001/03/03 10:01 [medline]
PHST- 2001/01/03 11:00 [entrez]
AID - tpj908 [pii]
AID - 10.1046/j.1365-313x.2000.00908.x [doi]
PST - ppublish
SO  - Plant J. 2000 Dec;24(6):713-23. doi: 10.1046/j.1365-313x.2000.00908.x.