Functional Analyses of c.2268dup in Thyroid Peroxidase Gene Associated with Goitrous Congenital Hypothyroidism
Figure 2
Agarose gel electrophoresis of reverse-transcription PCR (RT-PCR) products. (a) The RT-PCR products of exons 2 to 13 that include the exon-exon junctions. Abnormal spliced products were not detected in exons 2 to 12 based on the expected PCR products present (lanes 3 to 5). However, an unknown product of approximately 350 bp was detected for the exons 12/13 fragment in addition to the expected product of 300 bp. Lane 1: GeneRuler 100 bp DNA Ladder; lane 2: negative control; lane 3: exons 2 to 7, 671 bp; lane 4: exons 7 to 9, 803 bp; lane 5: exons 9 to 12, 499 bp and 328 bp; lane 6: exons 12 to 13, 300 bp and an unknown product of 350 bp. (b) The RT-PCR products of exons 7 to 9 and exons 13 to 17 that include the exon-exon junctions. Abnormal spliced products were not detected in exons 7 to 9 and exons 13 to 17 based on the expected PCR products present (lanes 3 to 6). Lane 1: GeneRuler 100 bp DNA Ladder; lane 2: negative control; lane 3: exons 13 to 15, 250 bp and 118 bp; lane 4: exons 15 to 17, 261 bp and 131 bp; lane 5: exons 7 to 8, 488 bp; lane 6: exons 8 to 9, 437 bp. (c) The RT-PCR products of exons 12 to 13 that include the exon-exon junction. An unknown product of approximately 350 bp and an expected product of 300 bp were detected in all thyroid tissue specimens from proband and III-1 with c.2268dup mutation. Lane 1: GeneRuler 100 bp DNA Ladder; lane 2: normal area of thyroid tissue of III-1; lane 3: lesion area of thyroid tissue of III-1; lane 4: normal area of thyroid tissue of proband; lane 5: lesion area of thyroid tissue of proband; lane 6: negative control. (d) The RT-PCR products of exons 12 to 13 that include the exon-exon junction. An expected product of 300 bp was detected in all c.2268dup-mutation free individuals.Lane 1: GeneRuler 100 bp DNA Ladder; lane 2: Control I; lane 3: Control II; lane 4: Control III.
