are desirable because of simple segregation patterns, among other reasons. As an example,
Figure 11.6 shows a plasmid used inAgrobacterium-mediated transformation. Figure 11.7
shows the T-DNA from right border (RB) to left border (LB) that would be transferred to
the plant genome usingAgrobacterium.Genomic DNA of the transgenic plant would
be digested with EcoR1, an enzyme that cuts relative to the left T-DNA border and
would cut into the plant genome, and the blot would be hybridized with the selectable
Figure 11.5.The downward Southern blot apparatus. Stacks of paper towel are topped with filter
paper, membrane, the gel, a filter paper wick, more filter paper, and a weight. DNA travels into the
membrane via wicking, capillary action and gravity.
Figure 11.6.Map of plasmid pJZ1. The map shows the location of Pro:SM:Term
(Promoter:Selectable Marker:Terminator), the Pro:GOI:Term (Promoter:Gene of Interest:
Terminator), and the ampicillin resistance gene (amp). Location of theEcoR1 andSac1 sites are
shown for the purpose of restriction enzyme digestion for Southern analysis. (LB¼left border of
T-DNA; RB¼right border of T-DNA.) The T-DNA between the left and right borders would be
transferred in anAgrobacterium-mediated transformation event. A particle bombardment vector
would not carry the LB and RB sequences.
282 TRANSGENIC PLANT ANALYSIS