Fig. 2.
Fig. 2. Analysis of normal and S.M.'s. / TAPASIN genes. (A) Genomic polymerase chain reaction (PCR) of a TAPASIN gene. PCR products were analyzed by 1.5% agarose gel electrophoresis. Odd lanes show 1B1-LCL (healthy control). Even lanes show SM-LCL. PCR primers used were as follows: exon 1-3; 5′-AGGAAGAGGAGGCTTCATGG-3′(forward), 5′-CCTTAGAATCTACCCACCCTT-3′(reverse); exons 4-7, 5′-TCATGCCCCTTCCCAACCC-3′(forward), 5′-ATAGGATGAGGAGATAAAGTGA-3′(reverse); exon 8, 5′-TTAAGAAGATGCAGGAGATACA-3′(forward), 5′-CACTCAGTGGAGAGAGATTG-3′(reverse); intron 3, 5′-CAGGCCTATACTTTCTTAAT-3′ (forward), 5′-ACCCCTTTCTTGTGCCTAGT-3′(reverse); intron 7, 5′-AACAATATAAGAAATTACAGGC-3′(forward), 5′-TGTATCTCCTGCATCTTCTTAA-3′(reverse). PCR products sizes are as follows: exons 1-3, 908 base pairs (bp); exons 4-7, 1529 bp; exon 8, 305 bp; intron 3, 1711 bp; intron 7, 1038 bp. (B) Schematic of normal and S.M.'s TAPASIN gene structure. Upper line shows normalTAPASIN gene; lower line, S.M.'s TAPASIN gene. Boxes indicate exons. PCR primer positions are indicated by arrowheads (exon primers) or arrows (intron primers; black arrows, intron 3; white arrows, intron 7, respectively). A 7.4-kilobase (kb) portion indicated by doted lines was deleted in S.M.'s TAPASIN gene. Scales of exons and introns are approximations. (C) Intron 3 structure of S.M.'s TAPASIN gene. Upper line shows normal tapasin intron 3 region; middle line, S.M.'s intron 3 region; lower line, normal tapasin intron 7 region. Arrows indicate putative breakpoints. Boxes indicate exons and Alu-repetitive sequences. In SM-tpn, open boxes correspond to the exon 3 to exon 4 region; shaded boxes correspond to the exon 7 to exon 8 region of normal TAPASIN gene. A hatched box (marked with arrows) indicates a putative recombination region. (D) Nucleotide sequence of putative recombination junction. Upper sequence shows normal tapasin intron 3 AluSp region; middle sequence, S.M.'s tapasin; lower sequence, normal tapasin intron 7 AluSx region. Arrows indicate differences in nucleotide sequence positions between S.M.'s and normal intron 3 or intron 7. Box indicates a putative recombination junction area.

Analysis of normal and S.M.'s

TAPASIN genes. (A) Genomic polymerase chain reaction (PCR) of a TAPASIN gene. PCR products were analyzed by 1.5% agarose gel electrophoresis. Odd lanes show 1B1-LCL (healthy control). Even lanes show SM-LCL. PCR primers used were as follows: exon 1-3; 5′-AGGAAGAGGAGGCTTCATGG-3′(forward), 5′-CCTTAGAATCTACCCACCCTT-3′(reverse); exons 4-7, 5′-TCATGCCCCTTCCCAACCC-3′(forward), 5′-ATAGGATGAGGAGATAAAGTGA-3′(reverse); exon 8, 5′-TTAAGAAGATGCAGGAGATACA-3′(forward), 5′-CACTCAGTGGAGAGAGATTG-3′(reverse); intron 3, 5′-CAGGCCTATACTTTCTTAAT-3′ (forward), 5′-ACCCCTTTCTTGTGCCTAGT-3′(reverse); intron 7, 5′-AACAATATAAGAAATTACAGGC-3′(forward), 5′-TGTATCTCCTGCATCTTCTTAA-3′(reverse). PCR products sizes are as follows: exons 1-3, 908 base pairs (bp); exons 4-7, 1529 bp; exon 8, 305 bp; intron 3, 1711 bp; intron 7, 1038 bp. (B) Schematic of normal and S.M.'s TAPASIN gene structure. Upper line shows normalTAPASIN gene; lower line, S.M.'s TAPASIN gene. Boxes indicate exons. PCR primer positions are indicated by arrowheads (exon primers) or arrows (intron primers; black arrows, intron 3; white arrows, intron 7, respectively). A 7.4-kilobase (kb) portion indicated by doted lines was deleted in S.M.'s TAPASIN gene. Scales of exons and introns are approximations. (C) Intron 3 structure of S.M.'s TAPASIN gene. Upper line shows normal tapasin intron 3 region; middle line, S.M.'s intron 3 region; lower line, normal tapasin intron 7 region. Arrows indicate putative breakpoints. Boxes indicate exons and Alu-repetitive sequences. In SM-tpn, open boxes correspond to the exon 3 to exon 4 region; shaded boxes correspond to the exon 7 to exon 8 region of normal TAPASIN gene. A hatched box (marked with arrows) indicates a putative recombination region. (D) Nucleotide sequence of putative recombination junction. Upper sequence shows normal tapasin intron 3 AluSp region; middle sequence, S.M.'s tapasin; lower sequence, normal tapasin intron 7 AluSx region. Arrows indicate differences in nucleotide sequence positions between S.M.'s and normal intron 3 or intron 7. Box indicates a putative recombination junction area.

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