Figure 4

T-DNA insertion sites of transgenic rice DR24 and MDH13. LB: the left border of T-DNA; RB: the right border of T-DNA; LSP1 and LSP2 are specific nested primers designed in the upstream of ltp promoter, MSP1 and MSP2 are specific nested primers designed downstream of ZmMDH gene, AP1 and Ap2 are specific nested primers designed in adapter, DJ-F and DJ-R are detection primers designed according to the right border sequence and right flanking sequence of T-DNA in DR24, and MJ-F and MJ-R are detection primers designed according to the left border sequence and left flanking sequence of T-DNA in MDH13, all the primers sequence used here are listed in Table 2. (a) T-DNA insertion site of DR24 is between 16789099 bp and 16789100 bp on Chromosome 9; (b) the T-DNA insertion site of DR24 is between 10844985 bp and 10844986 bp on Chromosome 9; (c) The boundary PCR detection, lane 1 is PCR band about 1.5 kb with detection primers DJ-F and DJ-R and lane 2 is PCR band about 4.3 kb with detection primers MJ-F and MJ-R. Sequencing from the two bands showed that the the insertion sites of DR24 and MDH13 transgenic lines to be correct.