[Amplification and analysis of T-DNA flanking sequences in transgenic rice].
Rice transformation mediated by Agrobacterium tumefaciens has technically matured to some extent, but the mechanics of T-DNA integration in transgenic rice remains largely unknown. Using thermal asymmetric interlaced PCR (TAIL-PCR), we analyzed the flanking sequences of T-DNAs in transgenic rice plants, in which the resistance gene for rice bacterial blight disease, Xa21, had been integrated stably. Sequence analysis of 24 fragments amplified by TAIL-PCR showed that of them 14 were rice genomic DNA, 9 contained vector backbone sequences, and one was a fragment of the exogenous gene Xa21. The characteristics of the 14 rice genomic DNA sequences at T-DNA integrated sites are significantly different from those of the reported rice genomic sequences into which exogenous genes were integrated through direct DNA transformation methods. T-DNA border sequences integrated in rice genome have similar features as those integrated in dicotyledonous genomes. In the backbone containing flanking sequences (37.5%, 9/24) vector backbones appeared in different types.
Fang
J
J
Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, China.
Zhai
W X
WX
Wang
W M
WM
Li
S W
SW
Zhu
L H
LH
D001483:Base Sequence; D004269:DNA, Bacterial / Q000737:chemistry*; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA