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

• Published in: African journal of biotechnology Vol. 10; no. 59; pp. 12614 - 12625
• Main Authors: Yang, L, Fu, F-L, Zhang, Z-Y, Zhou, S-F, She, Y-H, Li, W-C
• Format: Journal Article
• Language: English
• Published: 03.10.2011
• Subjects: Agrobacterium tumefaciens; Zea mays
• ISSN: 1684-5315; 1684-5315
• DOI: 10.5897/AJB11.728

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.