Molecular cloning of cDNAs encoding bioactive peptides by the sequence capture method

• Project/Area Number: 19570063
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Morphology/Structure
• Research Institution: Toho University
• Principal Investigator: IWAMURO Shawichi Toho University, 理学部, 准教授 (70221794)
• Co-Investigator(Kenkyū-buntansha): KIKUYAMA Sakae 早稲田大学, 教育・総合科学学術院, 名誉教授 (20063638) ; KOBAYASHI Tetsuya 埼玉大学, 理工学研究科, 准教授 (00195794)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2008: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2007: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 動物形態 / 両生類 / 抗菌ペプチド / 皮膚 / 遺伝子クローニング / ヒストンH2B / ファブリキウス嚢 / シュレーゲルアオガエル / 抗菌活性 / ハチ毒メリチン様ペプチド / ヒストンH4 / 細胞溶解性

Research Abstract

本研究では、両生類の皮膚から抗菌性のペプチドの単離を行なうとともに、その効率的な遺伝子クローニング法の開発を行なった。その結果、日本産の数種類のカエルから有用な抗菌ペプチド及びその配列を多数見つけ出すとともに、抗菌ペプチド遺伝子の発現には甲状腺ホルモンが重要であることを示した。また、ヒストンH2Bという本来は細胞の核にあるタンパク質がカエルの皮膚から得られていることに基づき、その抗菌メカニズムを明らかにした。

Research Products

• [Journal Article] Developmental and thyroid hormone induced expression of preprotemporin genes in the skin of Japanese mountain brown frog Rana ornativentris (2009)
  • Author(s): Ohnuma A, Conlon JM, Iwamuro S
  • Journal Title: Annals NY Acad Sci (in press)
• [Journal Article] Dermal cytolytic peptides and the system of innate immunity in anurans (2009)
  • Author(s): Conlon JM, Iwamuro S, King JD
  • Journal Title: Annals NY Acad Sci (in press)
• [Journal Article] Demonstration of the proopiomelaocortin signalling system in the primary immune organ, the bursa of Fabricius, of the quail (2009)
  • Author(s): Kobayashi T, Takaso A, Tanaka S, Amano M, Takahashi A, Iwamuro S, Machida, T
  • Journal Title: Annals NY Acad Sci (in press)
• [Journal Article] Developmental and thyroid hormone-induced expression of preprotemporin genes in the skin of Japanese mountain brown frog Rana ornativentris. (2009)
  • Author(s): Ohnuma A, Conlon JM, Iwamuro S
  • Journal Title: Annals of New York Academy of Sciences (im press)
  • Peer Reviewed
• [Journal Article] Dermal cytolytic peptides and the system of innate immunity in anurans. (2009)
  • Author(s): Conlon JM, Iwamuro S, King JD
  • Journal Title: Annals of New York Academy of Sciences (im press)
  • Peer Reviewed
• [Journal Article] Demonstration of the proopiomelaocortin signalling system in the primary immune organ, the bursa of Fabricius, of the quail. (2009)
  • Author(s): Kobayashi T, Takaso A, Tanaka S, Amano M, Takahashi A, Iwamuro S, Machida T
  • Journal Title: Annals of New York Academy of Sciences (im press)
  • Peer Reviewed
• [Journal Article] Potential roles of histones in host defense as antimicrobial agents (2008)
  • Author(s): Kawasaki H, Iwamuro S
  • Journal Title: Infectious Disorder-Drug Targets 8 Pages: 195-205
• [Journal Article] Antimicrobial action of histone H2B in Escherichia coli : evidence for membrane translocation and DNA-binding of a histone H2B fragment after proteolytic cleavage by outer membrane proteinase T (2008)
  • Author(s): Kawasaki H, Koyama T, Conlon JM, Yamakura F, Iwamuro S
  • Journal Title: Biochimie 90 Pages: 1693-1702
• [Journal Article] Molecular cloning of a cDNA encoding atypical antimicrobial and cytotoxic brevinin-1Ja from the skin of the Japanese brown frog, Rana japonica (2008)
  • Author(s): Koyama T, Iwamuro S
  • Journal Title: Zool Sci 25 Pages: 487-491
• [Journal Article] Characterization of a hemolytic protein, identified as histone H4, from the skin of the Japanese tree frog Hyla japonica(Hylidae) (2008)
  • Author(s): Kawasaki H, Iwamuro S, Goto Y, Nielsen Per F, Conlon JM
  • Journal Title: Comp Biochem Phys B 149 Pages: 120-125
• [Journal Article] Antimicrobial action of histone H2B in Escherichia coli : evidence for membrane translocation and DNA-binding of a histone H2B fragment after proteolytic cleavage by outer membrane proteinase T. (2008)
  • Author(s): Kawasaki H, Koyama T, Conlon JM, Yamakura F, Iwamuro S.
  • Journal Title: Biochimie 90 Pages: 1693-1702
  • Peer Reviewed
• [Journal Article] Potential roles of histones in host defence as antimicrobial agents. (2008)
  • Author(s): Kawasaki H, Iwamuro S.
  • Journal Title: Drug Targets Infectious Disorders 8 Pages: 195-205
  • Peer Reviewed
• [Journal Article] Characterization of a hemolytic protein, identified as histone H4, from the skin of the Japanese tree frog Hyla japonica(Hylidae). (2008)
  • Author(s): Kawasaki H, Iwamuro S, Goto Y, Nielsen PF, Conlon JM.
  • Journal Title: Comp Biochem Physiol B Biochem Mol Biol. 149 Pages: 120-125
  • Peer Reviewed
• [Journal Article] Molecular cloning of a cDNA encoding the atypical antimicrobial and cytotoxic brevinin-1Ja from the skin of Japanese brown frog Rana japonica. (2008)
  • Author(s): Koyama T, Iwamuro S.
  • Journal Title: Zoological Science (In press)
  • Peer Reviewed
• [Journal Article] Evidence that the genes encoding the melittin-related peptides in the skins of the Japanese frogs Rana sakuraii and Rana tagoi are not orthologous to bee venom melittin genes : developmental-and tissue-dependent gene expression (2007)
  • Author(s): Suzuki H, Conlon JM, Iwamuro, S
  • Journal Title: Peptides 28 Pages: 2061-2068
• [Journal Article] Cytolytic peptides belonging to the brevinin-1 and brevinin-2 families isolated from the skin of the Japanese brown frog, Rana dybowskii (2007)
  • Author(s): Conlon JM, Kokodziejek J, Nowotny N, Leprince J, Vaudry H, Coquet L, Jouenne T, Iwamuro S
  • Journal Title: Toxicon 50 Pages: 746-756
• [Journal Article] Expression of the estrogen receptor a gene in the anal fin of Japanese medaka, Oryzias latipes, by environmental concentrations of bisphenol A (2007)
  • Author(s): Hayashi H, Nishimoto A, Oshima N, Iwamuro S
  • Journal Title: J Toxicol Sci 32 Pages: 91-96
• [Journal Article] Antimicrobial peptides from the skin of the frog, Rana dybowskii implications for the phylogeny of the Japanese brown frogs. (2007)
  • Author(s): Conlon JM, Kolodziejek J, Nowotny N, Leprince J, Vaudry H, Coquet L, Jouenne T, Iwamuro, S.
  • Journal Title: Toxicon 50 Pages: 746-756
  • Peer Reviewed
• [Journal Article] Evidence that the genes encoding the melittin-related peptides in the skins of the Japanese frogs Rana sakuraii and Rana tagoi are not orthologous to bee venom melittin genes: developmental-and tissue-dependent gene expression. (2007)
  • Author(s): Suzuki H, Conlon JM, Iwamuro S.
  • Journal Title: Peptides 28 Pages: 2061-2068
  • Peer Reviewed
• [Presentation] 対馬産Rana属皮膚抗菌ペプチドの遺伝子クローニング (2009)
  • Author(s): 秋葉貴久、安藤麻理、Conlon JM、岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
  • Year and Date: 2009-03-20
• [Presentation] ウシガエルハーダー腺における抗菌ペプチド遺伝子の発現 (2009)
  • Author(s): 小西健司、蓮沼至、小林哲也、岡田令子、中野真樹、白間一彦、菊山榮、岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
  • Year and Date: 2009-03-20
• [Presentation] ゲノム情報から両生類抗菌ペプチド遺伝子の発現調節機構を探る (2009)
  • Author(s): 宮崎希、太枝志帆、竹山留美子、大沼彩、岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
  • Year and Date: 2009-03-20
• [Presentation] 無尾目両生類皮膚腺の形態学的解析 (2009)
  • Author(s): 丸橋佳織、岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
  • Year and Date: 2009-03-20
• [Presentation] 鳥類血球系及び免疫系からのCathelicidin遺伝子のクローニング (2009)
  • Author(s): 椿卓、小林哲也、稲嶺由羽、菊山榮、岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
  • Year and Date: 2009-03-20
• [Presentation] ウシガエルバーダー腺における抗菌ペプチド遺伝子の発現 (2009)
  • Author(s): 小西健司, 蓮沼至, 小林哲也, 岡田令子, 中野真樹, 自間一彦, 菊山榮, 岩室祥一
  • Organizer: 日本動物学会関東支部第61回大会
  • Place of Presentation: 埼玉大学
• [Presentation] ヒストンH2Bの抗大腸菌作用機序の解析 (2008)
  • Author(s): 岩室祥一、川崎広明、山倉文幸、Conlon J M.
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会合同大会(BMB2008)
  • Place of Presentation: 神戸国際展示場
  • Year and Date: 2008-12-10
• [Presentation] 両生類皮膚抗菌ペプチドの遺伝子発現とその甲状腺ホルモンによる関与 (2008)
  • Author(s): 岩室祥一、太枝志帆、鈴木啓恵、大沼彩
  • Organizer: 第33回日本比較内分泌学会大会
  • Place of Presentation: 広島大学
  • Year and Date: 2008-12-04
• [Presentation] ニホンアカガエル新規抗菌ペプチドBrevenin-1Ja (2008)
  • Author(s): 岩室祥一、小山卓己
  • Organizer: 第60回日本動物学会関東支部大会
  • Place of Presentation: 東京大学
  • Year and Date: 2008-03-22
• [Presentation] ニホンアカガエル抗菌ペプチドの遺伝子クローニングとその解析 (2008)
  • Author(s): 小山卓己、岩室祥一
  • Organizer: 第60回日本動物学会関東支部大会
  • Place of Presentation: 東京大学
  • Year and Date: 2008-03-22
• [Presentation] Molecular cloning and developmental expression of genes encoding antimicrobial peptide precursors in Rana ornativentris (2008)
  • Author(s): Iwamuro S, Oeda S, Miyazaki N, Conlon JM, Ohnuma A
  • Organizer: The 24th Conference of European Comparative Endocrinologists 2008, Genoa, Italy
  • Place of Presentation: Centro Congressi Magazzini del Cotone Centro Congressi Magazzini del Cotone
• [Presentation] Identification of Des-acetyl-alpha-melanocyte-stimulating hormone in the quail bursa of Fabricius (2008)
  • Author(s): Kobayashi T, Takaso A, Tanaka S, Amano M, Takahashi A, Iwamuro S, Machida T
  • Organizer: The 24th Conference of European Comparative Endocrinologists 2008, Genoa, Italy
  • Place of Presentation: Centro Congressi Magazzini del Cotone
• [Presentation] Molecular cloning and developmental expression of genes encoding antimicrobial peptide precursors in Ranaornativentris. (2008)
  • Author(s): Iwamuro s, Oeda S, Miyazaki N, Conlon J, Ohnuma A.
  • Organizer: The 24th Conference of European Comparative Endocrinologists 2008
  • Place of Presentation: Genoa, Italy
• [Presentation] Dermal cytolytic peptides and the system of innate immunity in amphibians. (2008)
  • Author(s): Conlon JM, Iwamuro S.
  • Organizer: The 24th Conference of European Comparative Endocrinologists 2008
  • Place of Presentation: Genoa, Italy
• [Presentation] Identification of Des-acetyl-alpha-melanocyte-stimulating hormone in the quail bursa of Fabricius. (2008)
  • Author(s): Kobayashi T, Takaso A, Tanaka S, Amano M, Takahashi A, Iwamuro S, Machida T.
  • Organizer: The 24th Conference of European Comparative Endocrinologists 2008
  • Place of Presentation: Genoa, Italy
• [Presentation] ヒストンH2Bの抗大腸菌作用機序の解析 (2008)
  • Author(s): 岩室祥一, 川崎広明, 山倉文幸, Conlon JM
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会合同大会(BMB2008)
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] 微生物を撃て! -カエルの抗菌ペプチド (2007)
  • Author(s): 岩室祥一
  • Organizer: 日本動物学会第78回大会シンポジウム「両生類はワンダーランド」
  • Place of Presentation: 弘前大学
  • Year and Date: 2007-09-22
• [Presentation] ヤマアカガエル抗菌ペプチドの遺伝子クローニングとそのステージ依存的な発現 (2007)
  • Author(s): 大沼彩、鈴木啓恵、Conlon, JM、岩室祥一
  • Organizer: 日本動物学会第78回大会
  • Place of Presentation: 弘前大学
  • Year and Date: 2007-09-20
• [Presentation] 組換え体ヒストンH2Bの抗大腸菌作用機構 (2007)
  • Author(s): 川崎広明、Conlon JM、小林哲也、岩室祥一
  • Organizer: 日本動物学会第78回大会
  • Place of Presentation: 弘前大学
  • Year and Date: 2007-09-20
• [Presentation] ニホンアカガエル由来抗菌ペプチドJaponicinの遺伝子クローニング及び局在解析 (2007)
  • Author(s): 小山卓己、Conlon JM、岩室祥一
  • Organizer: 日本動物学会第78回大会
  • Place of Presentation: 弘前大学
  • Year and Date: 2007-09-20
• [Presentation] アカガエル属抗菌ペプチドTemporinの遺伝子解析 (2007)
  • Author(s): 太枝志帆、大沼彩、竹山瑠美子、Conlon JM、岩室祥一
  • Organizer: 日本動物学会第78回大会
  • Place of Presentation: 弘前大学
  • Year and Date: 2007-09-20
• [Presentation] 両生類下垂体研究にまつわるささやかなセレンディピティー (2007)
  • Author(s): 菊山 榮
  • Organizer: 日本動物学会第78回大会シンポジウム「両生類はワンダーランド」
  • Place of Presentation: 弘前大学
• [Presentation] 微生物を撃て!-カエルの抗菌ペプチド (2007)
  • Author(s): 岩室祥一
  • Organizer: 日本動物学会第78回大会シンポジウム「両生類はワンダーランド」
  • Place of Presentation: 弘前大学