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Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation
Lox sites of the Cre/ lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon...
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Published in: | Cytology and genetics 2013-05, Vol.47 (3), p.145-155 |
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Main Authors: | , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Lox
sites of the Cre/
lox
recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of
lox
sites were tested for their effect upon bar gene expression in plants obtained via
Agrobacterium
-mediated and biolistic transformation methods.
Lox
-mediated expression of
bar
gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via
Agrobacterium
-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via
Agrobacterium
-mediated transformation was caused by activation of
bar
gene. The plasmid with promoterless
gus
gene together with the
lox
site adjacent to the RB was constructed and transferred to
Nicotiana tabacum
as well. Transgenic plants exhibited GUS activity and expression of
gus
gene was detected in plant leaves. Expression of
bar
gene from the vectors containing
lox
site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as
Beta vulgaris, Brassica napus, Lactuca sativa
and
Solanum tuberosum
. Our results demonstrate that the
lox
site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants. |
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ISSN: | 0095-4527 1934-9440 |
DOI: | 10.3103/S0095452713030079 |