Integration and characterization of T-DNA insertion in upland cotton
X. Yang, F. Li, X. Zhang, K. Liu, Q. Wang, C. Zhang, C. Liu, W. Zhu, G. Shan, C.K: Chin, W. Fanghttps://doi.org/10.17221/217/2012-CJGPBCitation:Yang X., Li F., Zhang X., Liu K., Wang Q., Zhang C., Liu C., Zhu W., Shan G., Chin C.K., Fang W. (2013): Integration and characterization of T-DNA insertion in upland cotton. Czech J. Genet. Plant Breed., 49: 51-57.
Copy numbers were evaluated by real-time quantitative PCR, and 149 junctions of T-DNA were isolated by thermal asymmetric interlaced PCR from 92 independent transgenic cotton lines transformed by Agrobacterium tumefaciens strain LBA4404. Real-time quantitative PCR results showed that 46% had integration of one or two T-DNA copies, 54% had three or more copies. Among 63 amplified products at LB junctions, 51% showed co-transformation of the vector backbone, 30% retained a portion of LB ranging from 3 to 23 bp, and 19% showed deletions ranging from 1 to 148 bp from the LB inner end. In contrast, all of the cleavage sites were located in the inner region of RB. The distribution of T-DNA insertions in upland cotton genome included coding sequences, transposons, plastid-derived sequences and microsatellites.
cotton (Gossypium hirsutum L.); deletion of border; genetic transformation; transgene copy; vector integration