TY - JOUR
T1 - Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization
AU - Pedersen, C.
AU - Zimny, J.
AU - Becker, D.
AU - Jähne-Gärtner, A.
AU - Lörz, H.
PY - 1997/5/1
Y1 - 1997/5/1
N2 - Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.
AB - Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.
KW - Chromosomal localization
KW - FISH
KW - Integration pattern
KW - Transgenic barley
KW - Wheat and triticale
UR - http://www.scopus.com/inward/record.url?scp=0030955688&partnerID=8YFLogxK
U2 - 10.1007/s001220050474
DO - 10.1007/s001220050474
M3 - Journal article
AN - SCOPUS:0030955688
SN - 0040-5752
VL - 94
SP - 749
EP - 757
JO - Theoretical And Applied Genetics
JF - Theoretical And Applied Genetics
IS - 6-7
ER -