Figure 3. Construction of mutant plasmids. A: PCR products of the recombinant mutant TGFBI-T538P and TGFBI-R555W genes. M: 1 kb marker (Fermentas, Burlington, Canada),
lane 1: TGFBI-T538P, lane 2: TGFBI-R555W. B: Constructed pcTGFBI-WT/T538P/R555W-myc plasmids were double digested into two bands approximately 2 kb and 5.5 kb, respectively,
using Xho I and Hind III. M: 1 kb marker, lane 1: pcTGFBI-WT-myc, lane 2–lane 3: pcTGFBI-T538P-myc, lane 4–lane 5: pcTGFBI-R555W-myc.
C: Right: Partial sequence of the pcTGFBI-T538P plasmid showing the A to C change at position c.1613 (the forward strand is
shown) resulting in the p.Thr538Pro mutation (underlined). Left: Partial sequence of the pcTGFBI-R555W plasmid showing the
C to T mutation at position c.1663 (the forward strand is shown) resulting in the p.Arg555Trp mutation (underlined). The arrow
points toward the position of the mutations.