Purpose: To compare heteroduplex analysis with SSCP and to develop a simple and effective method for mutational screening of RB gene.Materials and Methods: Leukocyte DNA was prepared from 12 unrelated Japanese patient...Purpose: To compare heteroduplex analysis with SSCP and to develop a simple and effective method for mutational screening of RB gene.Materials and Methods: Leukocyte DNA was prepared from 12 unrelated Japanese patients with hereditary retinoblstoma. PCR combined with simultaneous nonisotopic heteroduplex and SSCP analysis was used to screen leukocyte DNA for such mutations, exon-by-exon, without the use of restriction endonu-clease digestion. PCR was conducted using 28 pairs of primers flanking all 27 ex-ons and the promoter region of the RB gene, with PCR products ranging from 159bp to 326bp. Mutations were identified by sequencing.Results: Heterozygous germline mutations were detected in 8 of 12 Japanese patients. The mutations were identified by sequencing as follows; G→C/acceptor of exon 11, T insertion/codon 389, C→T/codon 455, 33bp insertion/codon 455 (C GA), G→T/codon 533, C→T/codon 579, C deletion/codon 674, and C→T/ codon 787.Conclusion: Our results suggest that small RB gene mutations展开更多
Purpose: To disclose the structure of visual pigment gene for a protanopia with specific variation.Methods: Exon 5 fragments of the red andgreen visual pigment genes from the protanopia with specific varnation as well...Purpose: To disclose the structure of visual pigment gene for a protanopia with specific variation.Methods: Exon 5 fragments of the red andgreen visual pigment genes from the protanopia with specific varnation as well as controls were amplified by poly-merase chain reaction (PCR). The PCR products were put through heteroduplex-SSCP analysis and PCR-RFLP (restriction fragement length polymorphism) analysis to clarify the specific variation. The specific variation of the exon 5 DNA fragment from the protanopia was identified by sequencing.Results: A novel 5’green-3’red hybrid gene fragment without the normal red and green visual pigment gene was discovered in the protanopia. He should only have a single visual pigment gene, 5’green-3’red hybrid gene, on his X chromosome. The fusion point is between codon 285 and codon 296 in exon 5. Conclusion : Unequal intragenic recombination may occur in exon 5 as well as its upstream. A 5’green-3’red hybrid gene may present independently on the X chromosome without展开更多
Retinobiastoma is a highly malignant intraocular tumor of children that requires accurate diagnosis to prompt treatment. This article reviewed clinical, pathological and follow-up data on 1 147 cases of retinobiastoma...Retinobiastoma is a highly malignant intraocular tumor of children that requires accurate diagnosis to prompt treatment. This article reviewed clinical, pathological and follow-up data on 1 147 cases of retinobiastoma registered in Japan from 1975 to 1982. It is obvious that the prognosis of children with retinobiastoma has improved remarkably in recent years. The current advances in the management of the retinobiastoma were discussed.展开更多
文摘Purpose: To compare heteroduplex analysis with SSCP and to develop a simple and effective method for mutational screening of RB gene.Materials and Methods: Leukocyte DNA was prepared from 12 unrelated Japanese patients with hereditary retinoblstoma. PCR combined with simultaneous nonisotopic heteroduplex and SSCP analysis was used to screen leukocyte DNA for such mutations, exon-by-exon, without the use of restriction endonu-clease digestion. PCR was conducted using 28 pairs of primers flanking all 27 ex-ons and the promoter region of the RB gene, with PCR products ranging from 159bp to 326bp. Mutations were identified by sequencing.Results: Heterozygous germline mutations were detected in 8 of 12 Japanese patients. The mutations were identified by sequencing as follows; G→C/acceptor of exon 11, T insertion/codon 389, C→T/codon 455, 33bp insertion/codon 455 (C GA), G→T/codon 533, C→T/codon 579, C deletion/codon 674, and C→T/ codon 787.Conclusion: Our results suggest that small RB gene mutations
文摘Purpose: To disclose the structure of visual pigment gene for a protanopia with specific variation.Methods: Exon 5 fragments of the red andgreen visual pigment genes from the protanopia with specific varnation as well as controls were amplified by poly-merase chain reaction (PCR). The PCR products were put through heteroduplex-SSCP analysis and PCR-RFLP (restriction fragement length polymorphism) analysis to clarify the specific variation. The specific variation of the exon 5 DNA fragment from the protanopia was identified by sequencing.Results: A novel 5’green-3’red hybrid gene fragment without the normal red and green visual pigment gene was discovered in the protanopia. He should only have a single visual pigment gene, 5’green-3’red hybrid gene, on his X chromosome. The fusion point is between codon 285 and codon 296 in exon 5. Conclusion : Unequal intragenic recombination may occur in exon 5 as well as its upstream. A 5’green-3’red hybrid gene may present independently on the X chromosome without
文摘Retinobiastoma is a highly malignant intraocular tumor of children that requires accurate diagnosis to prompt treatment. This article reviewed clinical, pathological and follow-up data on 1 147 cases of retinobiastoma registered in Japan from 1975 to 1982. It is obvious that the prognosis of children with retinobiastoma has improved remarkably in recent years. The current advances in the management of the retinobiastoma were discussed.
