PMID- 11319036
OWN - NLM
STAT- MEDLINE
DCOM- 20011018
LR  - 20240109
IS  - 0960-7412 (Print)
IS  - 0960-7412 (Linking)
VI  - 25
IP  - 6
DP  - 2001 Mar
TI  - Localization of single-copy T-DNA insertion in transgenic shallots (Allium cepa) 
      by using ultra-sensitive FISH with tyramide signal amplification.
PG  - 699-707
AB  - The sensitivity of fluorescence in situ hybridization (FISH) for mapping plant 
      chromosomes of single-copy DNA sequences is limited. We have adapted for plant 
      cytogenetics a new signal-amplification method termed tyramide-FISH (Tyr-FISH). 
      Until present this technique has only been applied to human chromosomes. The 
      method is based on enzymatic deposition of fluorochrome-conjugated tyramide. With 
      Tyr-FISH it was possible to detect target T-DNA sequences on plant metaphase 
      chromosomes as small as 710 bp without using a cooled CCD camera. Short detection 
      time and high sensitivity, in combination with a low background, make the 
      Tyr-FISH method very suitable for routine application in plant cytogenetic 
      research. With Tyr-FISH we analysed the position of T-DNA inserts in transgenic 
      shallots. We found that the inserts were preferentially located in the distal 
      region of metaphase chromosomes. Sequential fluorescence in situ hybridization 
      with a 375 bp satellite sequence suggested that a specific T-DNA insert was 
      located within the satellite sequence hybridization region on a metaphase 
      chromosome. Analysis of less-condensed prophase and interphase chromosomes 
      revealed that the T-DNA was integrated outside the satellite DNA-hybridization 
      region in a more proximal euchromatin region.
FAU - Khrustaleva, L I
AU  - Khrustaleva LI
AD  - Plant Research International, Wageningen University and Research Center, PO Box 
      16, 6700 AA Wageningen, The Netherlands.
FAU - Kik, C
AU  - Kik C
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
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Bacterial)
RN  - 0 (Genetic Markers)
RN  - 0 (T-DNA)
RN  - X8ZC7V0OX3 (Tyramine)
SB  - IM
MH  - Chromosome Mapping/*methods
MH  - Chromosomes/genetics
MH  - DNA Probes
MH  - DNA, Bacterial/*genetics/*isolation & purification
MH  - *Genes, Plant
MH  - Genetic Markers
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Metaphase
MH  - Onions/*genetics/ultrastructure
MH  - Plants, Genetically Modified
MH  - *Transgenes
MH  - Tyramine/analogs & derivatives
EDAT- 2001/04/25 10:00
MHDA- 2001/10/19 10:01
CRDT- 2001/04/25 10:00
PHST- 2001/04/25 10:00 [pubmed]
PHST- 2001/10/19 10:01 [medline]
PHST- 2001/04/25 10:00 [entrez]
AID - tpj995 [pii]
AID - 10.1046/j.1365-313x.2001.00995.x [doi]
PST - ppublish
SO  - Plant J. 2001 Mar;25(6):699-707. doi: 10.1046/j.1365-313x.2001.00995.x.