1996 年度実績報告書

ヒトゲノム構造解析

研究課題

研究課題/領域番号	08283108
研究種目	重点領域研究
研究機関	国立がんセンター
研究代表者	大木操国立がんセンター, 研究所・放射線研究部, 部長 (00158792)
研究分担者	戸田達史東京大学, 医科学研究所, 助教授 (30262025) 辻省次新潟大学, 脳研究所, 教授 (70150612) 猪子英俊東海大学, 医学部, 教授 (10101932) 清水信義慶應義塾大学, 医学部, 教授 (50162706) 榊佳之東京大学, 医科学研究所, 教授 (10112327)
キーワード	11番染色体物理地図 / ダウン症必須領域 / シークエンス解析 / 免疫グロブリン遺伝子 / HLA遺伝子群 / SCA2遺伝子 / 福山型筋ジストロフィー / 改良型SSCP法
研究概要	11番染色体長腕全域のNotI制限酵素地図をリンキングクローン法にて作成し完成した。全長83メガ塩基に達し、制限酵素地図としては最長のものである。長領域のシークエンス解析を行い、以下の成果を得た。1)ヒト21番染色体のダウン症必須領域1.4Mbのシークエンスを決定し、エキソン予測プログラム及び相同性検索プログラムを用いて遺伝子の検索を行った。更に、真の遺伝子である可能性の高いものについてcDNAクローニングを行い、遺伝子の全構造を決定した。その結果7個の遺伝子が見つかり、そのうち4個は新規遺伝子であった。2)免疫グロブリンλ遺伝子、及びH鎖遺伝子領域、各lMbの塩基配列を決めた。3)6番染色体HLA遺伝子群のクラスI領域計776kbの塩基配列の決定を行い、新発現遺伝子13個、新偽遺伝子6個を含む計38個の遺伝子を同定した。また、本領域に298個のマイクロサテライトを見出したが、これらのうちの1つのトリプレットリピート配列がMICA遺伝子のコーディング領域に存在し、その1つの多型(6回繰り返し)がベ-チエット病と強い相関を示すことを見つけた。遺伝性神経性疾患に共通する発症機構として注目されているトリプレットリピート病について、病的に伸長したトリプレットリピートを効率よく検出できる方法(direct identification of repeat expansion and cloning technique,DIRECT)を開発し、脊髄小脳変性症2型(SCA2)の原因遺伝子を同定した。福山型筋ジストロフィー原因遺伝子の解析はD9S2107を含む100kb以下の領域に限定した。がん抑制遺伝子の単離をLOHの解析を中心に進め、乳がん等で詳細な共通欠失領域(3p21)をみつけた。ゲノム解析技術では、5′Cap構造に着目し、完全長cDNAを選択的に捕捉する方法論を開発した。またSSCP法の改良を行い、800bpまでのDNA断片中の変異を効率よく検出する系を確立した。

研究成果

(32件)

すべてその他

すべて文献書誌 (32件)

[文献書誌] Ohira.M.: "Identification of a novel gene containing the tetratricopeptide repeat domain from the Down syndrome region of chromosome 21." DNA Res.3. 65-74 (1996)
[文献書誌] Arai.Y.: "A Yeast artificial chromosome contig and Not I restriction map that spans the tumor suppressor gene (s) locus.11q22.2・23.3." Genomics. 35. 196-206 (1996)
[文献書誌] Morohoshi.F.: "Cloning and mapping of a human RBP56 gene encoding a putative RNA binding protein similar to FUS/TLS and EWS protein." Genomics. 38. 51-57 (1996)
[文献書誌] Naruse.K.: "A YAC contig of the human CC chemokine gene clusterd on chromosome 17q11.2." Genomics. 34. 236-240 (1996)
[文献書誌] Tsukahara.F.: "Identification and cloning of a novel cDNA belonging to tetratricopeptide repeat gene family from Down syndrome-critical region 22q22.2." J.Biochem.120. 820-827 (1996)
[文献書誌] Shibata.A.: "Identification of cis-acting elements involved in an alternative splicing of the amyloid precursor protein (APP) gene." Gene. 175. 203-208 (1996)
[文献書誌] Hattori,M.: "Novel Method for making nested deletions and its application for sequencing of a 300kb APP gene locus." Nucleic Acid Res.in press.
[文献書誌] Kuro,J.: "Localization of 16 exons to a 450-kb region involved in the autoimmune polyglandular disease type I (Apeced) on human chromosome 21q22.3." DNA Res.in press.
[文献書誌] Kawasaki,K.: "One megabase sequence analysis of the human immunoglobulin λ gene locus." Genome Res.in press.
[文献書誌] Nagamine,K.: "Isolation of cDNA for a novel human protein KNP-I that is homologous to the E.coli SCRP-27A protein from the autoimmunune polyglandular disease type I (APCED) region of chromosome 21q22.3." Biochem.Biopys.Rse.Commun.225. 608-616 (1996)
[文献書誌] Ueda,Y.: "Tumor-specific rearrangements of the immunoglobulin heavy-chain gene in B-cell Non-Hodgkin's lymphoma detected by in situ hybridization." Blood. 87. 292-298 (1996)
[文献書誌] Fukugawa,T.: "Human pseudoautosomal boundary-like sequences (PABLs) ; core sequence,expression,and evolution." Human Molecular Genetics. 5. 23-32 (1996)
[文献書誌] Ishihara,M.: "Genetic polymorphisms of the MHC-encoded antigen processing gene (TAP,LMP) in sarcoidosis." Hum.lmmunl.45. 105-110 (1996)
[文献書誌] Kasahara,M.: "Chromosomal localization of the proteosome Z subunit gene reveals an ancient chromosomal duplication involving the major histocompatibility complex." Proc.Natl.Acad. sci,.USA. 93. 9096-9101 (1996)
[文献書誌] Mizuki,N.: "Triplet repeat polymorphism in the transmembrane region of the MICA gene : A strong association of six GCT repetitions with Behcet disease." Prec.Natl.Acad.Sci,.USA. 94. 1298-1303 (1997)
[文献書誌] Takano,H.: "Somatic mosaicism of expanded CAG repeats in brains of patients with dentatorubral-pallidoluysianatrophy : Cellular population-dependent dynamics of mitotic instability." Am.J.Hum.Genet.58. 1212-1222 (1996)
[文献書誌] Igarashi,S.: "Intergenerational instability of the CAG repeat of the gene for Machado-Joseph disease (MIDI) is affected by the genotype of the normal chromosome : implications for the molecular mechanisms of the instability of the CAG repeat." Hum.Mol.Genet.5. 923-932 (1996)
[文献書誌] Sanpei,K.: "Identification of the spinocerebellar ataxia type 2 gene using a derect identification of repeat expansion and cloning technique,DIRECT." Nature Genet.14. 277-284 (1996)
[文献書誌] Toda,T.: "Linkage-disequilibrium mapping narrows the Fukuyama-type congenital muscular dystrophy (FCMD) candidate regin to<100kb." Am.J.Hum.Genet.59. 1313-1320 (1996)
[文献書誌] Miyaka,M.: "YAC and cosmid contigs encompassing the Fukuyama-type congenital muscular dystrophy (FCMD) candidate region on 9q31." Genomics. in press.
[文献書誌] Ohta,T.: "Isolation of a cosmid clone corresponding to a region of the inv (21) breakpoint in a patient with transient abormal myelopoiesis." Am.J.Hum.Genet.58. 544-550 (1996)
[文献書誌] Iida,A.: "Localization of a breast cancer tumor-suppressor gene to a 3-cM interval within chromosomal region 16q22." Brit.J.Cancer. 75. 264-267 (1997)
[文献書誌] Suzuki,H.: "Three distinct commonly deleted regions of chromosome arm 16q in human primary and metastatic prostate cancers." Genes,Chrom.Cancer. 17. 225-233 (1996)
[文献書誌] Imai,T.: "Identification and characterization of a new gene physically linked to ATM gene." Genome Res.6. 439-447 (1996)
[文献書誌] Caminci,P.: "High-efficiency full-length cDNA cloning by biotinylated CAP trapper." Genomics. 37. 327-336 (1996)
[文献書誌] Yoshikawa,H.: "Chromosome assignment of human genomic Not I restriction fragments in a two-dimensional electrophoresis profile." Genomics. 31. 28-35 (1996)
[文献書誌] Ichikawa,D.: "Analysis of numerical aberrations in specific chromosomes by fluorescence in situ hybridization (FISH) as a diagnostic tool in breast cancer." Cancer. 77. 2064-2069 (1996)
[文献書誌] Kumura,T.: "The brain finger protein gene (ZNF197),a mumber of the RING finger family,maps within the Smith-Magenis syndrome region in 17p11.2." Am.J.Med.Genet.in press.
[文献書誌] Inazuka,M.: "One-tube post-PCR fluorescent labeling of DNA fragments." Genome Res.6. 551-557 (1996)
[文献書誌] Kanbara,H.: "Fragment walking for long DNA sequencing by using a library as small as 16 primers." Gene. 176. 231-235 (1996)
[文献書誌] Matsuda.F.: "Organization of the Human lmmunoglobulin Heavy-Chain Locus." Academic Press, 1-29 (1996)
[文献書誌] Hayashi.K.: ""PCR・SSCP".in "Laboratory Protocol for Mutation Detection"" Oxford University Press, 14-22 (1996)

1996 年度 実績報告書

ヒトゲノム構造解析

研究代表者

大木 操 国立がんセンター, 研究所・放射線研究部, 部長 (00158792)

研究成果

[文献書誌] Ohira.M.: "Identification of a novel gene containing the tetratricopeptide repeat domain from the Down syndrome region of chromosome 21." DNA Res.3. 65-74 (1996)

[文献書誌] Arai.Y.: "A Yeast artificial chromosome contig and Not I restriction map that spans the tumor suppressor gene (s) locus.11q22.2・23.3." Genomics. 35. 196-206 (1996)

[文献書誌] Morohoshi.F.: "Cloning and mapping of a human RBP56 gene encoding a putative RNA binding protein similar to FUS/TLS and EWS protein." Genomics. 38. 51-57 (1996)

[文献書誌] Naruse.K.: "A YAC contig of the human CC chemokine gene clusterd on chromosome 17q11.2." Genomics. 34. 236-240 (1996)

[文献書誌] Tsukahara.F.: "Identification and cloning of a novel cDNA belonging to tetratricopeptide repeat gene family from Down syndrome-critical region 22q22.2." J.Biochem.120. 820-827 (1996)

[文献書誌] Shibata.A.: "Identification of cis-acting elements involved in an alternative splicing of the amyloid precursor protein (APP) gene." Gene. 175. 203-208 (1996)

[文献書誌] Hattori,M.: "Novel Method for making nested deletions and its application for sequencing of a 300kb APP gene locus." Nucleic Acid Res.in press.

[文献書誌] Kuro,J.: "Localization of 16 exons to a 450-kb region involved in the autoimmune polyglandular disease type I (Apeced) on human chromosome 21q22.3." DNA Res.in press.

[文献書誌] Kawasaki,K.: "One megabase sequence analysis of the human immunoglobulin λ gene locus." Genome Res.in press.

[文献書誌] Nagamine,K.: "Isolation of cDNA for a novel human protein KNP-I that is homologous to the E.coli SCRP-27A protein from the autoimmunune polyglandular disease type I (APCED) region of chromosome 21q22.3." Biochem.Biopys.Rse.Commun.225. 608-616 (1996)

[文献書誌] Ueda,Y.: "Tumor-specific rearrangements of the immunoglobulin heavy-chain gene in B-cell Non-Hodgkin's lymphoma detected by in situ hybridization." Blood. 87. 292-298 (1996)

[文献書誌] Fukugawa,T.: "Human pseudoautosomal boundary-like sequences (PABLs) ; core sequence,expression,and evolution." Human Molecular Genetics. 5. 23-32 (1996)

[文献書誌] Ishihara,M.: "Genetic polymorphisms of the MHC-encoded antigen processing gene (TAP,LMP) in sarcoidosis." Hum.lmmunl.45. 105-110 (1996)

[文献書誌] Kasahara,M.: "Chromosomal localization of the proteosome Z subunit gene reveals an ancient chromosomal duplication involving the major histocompatibility complex." Proc.Natl.Acad. sci,.USA. 93. 9096-9101 (1996)

[文献書誌] Mizuki,N.: "Triplet repeat polymorphism in the transmembrane region of the MICA gene : A strong association of six GCT repetitions with Behcet disease." Prec.Natl.Acad.Sci,.USA. 94. 1298-1303 (1997)

[文献書誌] Takano,H.: "Somatic mosaicism of expanded CAG repeats in brains of patients with dentatorubral-pallidoluysianatrophy : Cellular population-dependent dynamics of mitotic instability." Am.J.Hum.Genet.58. 1212-1222 (1996)

[文献書誌] Igarashi,S.: "Intergenerational instability of the CAG repeat of the gene for Machado-Joseph disease (MIDI) is affected by the genotype of the normal chromosome : implications for the molecular mechanisms of the instability of the CAG repeat." Hum.Mol.Genet.5. 923-932 (1996)

[文献書誌] Sanpei,K.: "Identification of the spinocerebellar ataxia type 2 gene using a derect identification of repeat expansion and cloning technique,DIRECT." Nature Genet.14. 277-284 (1996)

[文献書誌] Toda,T.: "Linkage-disequilibrium mapping narrows the Fukuyama-type congenital muscular dystrophy (FCMD) candidate regin to<100kb." Am.J.Hum.Genet.59. 1313-1320 (1996)

[文献書誌] Miyaka,M.: "YAC and cosmid contigs encompassing the Fukuyama-type congenital muscular dystrophy (FCMD) candidate region on 9q31." Genomics. in press.

[文献書誌] Ohta,T.: "Isolation of a cosmid clone corresponding to a region of the inv (21) breakpoint in a patient with transient abormal myelopoiesis." Am.J.Hum.Genet.58. 544-550 (1996)

[文献書誌] Iida,A.: "Localization of a breast cancer tumor-suppressor gene to a 3-cM interval within chromosomal region 16q22." Brit.J.Cancer. 75. 264-267 (1997)

[文献書誌] Suzuki,H.: "Three distinct commonly deleted regions of chromosome arm 16q in human primary and metastatic prostate cancers." Genes,Chrom.Cancer. 17. 225-233 (1996)

[文献書誌] Imai,T.: "Identification and characterization of a new gene physically linked to ATM gene." Genome Res.6. 439-447 (1996)

[文献書誌] Caminci,P.: "High-efficiency full-length cDNA cloning by biotinylated CAP trapper." Genomics. 37. 327-336 (1996)

[文献書誌] Yoshikawa,H.: "Chromosome assignment of human genomic Not I restriction fragments in a two-dimensional electrophoresis profile." Genomics. 31. 28-35 (1996)

[文献書誌] Ichikawa,D.: "Analysis of numerical aberrations in specific chromosomes by fluorescence in situ hybridization (FISH) as a diagnostic tool in breast cancer." Cancer. 77. 2064-2069 (1996)

[文献書誌] Kumura,T.: "The brain finger protein gene (ZNF197),a mumber of the RING finger family,maps within the Smith-Magenis syndrome region in 17p11.2." Am.J.Med.Genet.in press.

[文献書誌] Inazuka,M.: "One-tube post-PCR fluorescent labeling of DNA fragments." Genome Res.6. 551-557 (1996)

[文献書誌] Kanbara,H.: "Fragment walking for long DNA sequencing by using a library as small as 16 primers." Gene. 176. 231-235 (1996)

[文献書誌] Matsuda.F.: "Organization of the Human lmmunoglobulin Heavy-Chain Locus." Academic Press, 1-29 (1996)

[文献書誌] Hayashi.K.: ""PCR・SSCP".in "Laboratory Protocol for Mutation Detection"" Oxford University Press, 14-22 (1996)

1996 年度実績報告書

大木操国立がんセンター, 研究所・放射線研究部, 部長 (00158792)