Amplification of Tail-PCR in Amplifying Flanking Sequence of T-DNA in Torenia*

Abstract

Torenia (Torenia fournieri Lind.) is an important flowering plant for scientific research with ornamental values. The flanking sequence of T-DNA insertion site was successfully amplif ied using TAIL-PCR in torenia,and the length of the amplified products ranged about from 200 bp to 2 000 bp and most of them were about 400 bp and 800 bp. Among them,36% had homologous plant sequences. By BLAST analysis in NCBI,the predicted protein encoded by the flanking sequence of T-DNA insertion site was determined and seven gene sequences were submitted to GenBank. In addition,the rule was analyzed for T-DNA integrated into plant genome,and it suggested that the T-DNA integration occurred in the T-DNA region 15～18 bp from right border (RB) and 234 bp from outside of the RB,and the percentages of total fragments were 47.62% and 38.1% respectively. These provide the guarantee in experimental techniques for gene cloning and functional study of torenia by T-DNA tagging. Fig 2,Tab 2,Ref 24