T-DNA integration patterns in transgenic maize lines mediated by Agrobacterium tumefaciens

Journal: African Journal of Biotechnology, 2011，10(59):12614-12625

Author: Lin Yang, Feng-Ling Fu, Zhi-Yong Zhang, Shu-Feng Zhou, Yue-Hui She, Wan-Chen Li

Abstract:

To explore transfer deoxyribonucleic acid (T-DNA) integration patterns in the maize genome, we improved the protocol of thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR), and amplified the flanking sequences around T-DNA integration sites from 70 independent transgenic maize lines mediated by Agrobacterium tumefaciens. Out of 64 specific amplified fragments, 32 and 9 are homologous to the sequences of the maize genome and the expression plasmid, respectively. For 26 of them, a filler sequence was found flanking the cleavage sites. These results demonstrate that cleavage occurs not only during the T-DNA borders but also inside or outside the borders. The border sequences and some inside sequences can be deleted, and filler sequences can be inserted. Illegitimate recombination is a major pattern of T-DNA integration, while some hot spots and preference are present on maize chromosomes.

Link: http://www.academicjournals.org/ajb/full%20text/2011/3Oct/Yang%20et%20al.htm