PMID- 12582875
OWN - NLM
STAT- MEDLINE
DCOM- 20030317
LR  - 20080215
IS  - 0040-5752 (Print)
IS  - 0040-5752 (Linking)
VI  - 106
IP  - 1
DP  - 2002 Dec
TI  - Use of fluorescence in situ hybridization for gross mapping of transgenes and 
      screening for homozygous plants in transgenic barley ( Hordeum vulgare L.).
PG  - 92-100
AB  - Introduced transgenes, uidA, sgfp (S65T) and/or bar, were localized using 
      fluorescence in situ hybridization (FISH) on metaphase chromosomes of transgenic 
      barley produced by microparticle bombardment of immature embryos. Of the 19 
      independent transgenic lines (eight diploid and 11 tetraploid), nine had uidA and 
      ten had s gfp (S65T). All lines tested had three or more copies of the transgenes 
      and 18 out of 19 lines had visibly different integration sites. At a gross level, 
      it appeared that no preferential integration sites of foreign DNA among 
      chromosomes were present in the lines tested; however, a distal preference for 
      transgene integration was observed within the chromosome. In diploid T0 plants 
      that gave a 3:1 segregation ratio of transgene expression in the T1, only single 
      integration sites were detected on one of the homologous chromosomes. Homozygous 
      diploid plants had doublet signals on a pair of homologous chromosomes. All 
      tetraploid T0 plants that gave a 3:1 segregation ratio in the T1 generation had 
      only a single integration site on one of the homologous chromosomes. In contrast, 
      the single tetraploid T0 plant with a 35:1 segregation ratio in the T1 generation 
      had doublet signals on a pair of homologous chromosomes. In the one tetraploid T0 
      line, which had a homozygote-like segregation ratio (45:0), there were doublet 
      signals at two loci on separate chromosomes. We conclude that the application of 
      FISH for analysis of transgenic plants is useful for the gross localization of 
      transgene(s) and for early screening of homozygous plants.
FAU - Choi, H W
AU  - Choi HW
AD  - Department of Plant and Microbial Biology, University of California, Berkeley, CA 
      94720, USA.
FAU - Lemaux, P G
AU  - Lemaux PG
FAU - Cho, M-J
AU  - Cho MJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20020718
PL  - Germany
TA  - Theor Appl Genet
JT  - TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik
JID - 0145600
RN  - EC 2.3.1.- (Acetyltransferases)
RN  - EC 2.3.1.- (phosphinothricin N-acetyltransferase)
SB  - IM
MH  - Acetyltransferases/genetics
MH  - *Chromosome Mapping
MH  - Gene Dosage
MH  - Gene Expression
MH  - Homozygote
MH  - Hordeum/*genetics
MH  - *In Situ Hybridization, Fluorescence
MH  - Phenotype
MH  - Plants, Genetically Modified
MH  - Polyploidy
MH  - *Transgenes
EDAT- 2003/02/13 04:00
MHDA- 2003/03/18 04:00
CRDT- 2003/02/13 04:00
PHST- 2001/11/14 00:00 [received]
PHST- 2002/04/18 00:00 [accepted]
PHST- 2003/02/13 04:00 [pubmed]
PHST- 2003/03/18 04:00 [medline]
PHST- 2003/02/13 04:00 [entrez]
AID - 10.1007/s00122-002-0997-y [doi]
PST - ppublish
SO  - Theor Appl Genet. 2002 Dec;106(1):92-100. doi: 10.1007/s00122-002-0997-y. Epub 
      2002 Jul 18.