Molecular cloning and expression analysis of acute phase reactants in bony fish.

硬骨鱼急性期反应物的分子克隆和表达分析。

• 批准号: 11460098
• 负责人: YANO Tomoki
• 金额: $ 8.83万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460098/
• 关键词: FISH; IMMUNOLOGY; ACUTE PHASE REACTANTS; CLONING; RT-PCR; CARP; TROUT; サイトカイン; 魚類; 遺伝子; B因子; 発現解析

Complementary DNAs encoding acute phase reactants in the common carp and rainbow trout were cloned by means of the suppression subtractive hybridization. Sodium alginate, turpentaine oil, scleroglucan, and formaline-killed Vibrio anguillarum were used as stimulants. As a result, about 50 genes including acute phase protein known in mammals, such as pentraxin and serum amyloid A, as well as several chemokines, chemokine-receptors, a novel C-type lectin, a novel complement factor B isotypes, glia-maturation factor, IL-1beta, and pre-B cell enhancing factor were identified from carp. On the other hand, 36 genes including LECT2 chemotaxin, complement B/C2, a C-type lectin, fibrinogen, haptoglobin, hemopexine, CD59, lysozyme, viterogenin, precelebrin, Toll-like receptors, transferrin, serum amyloid A, C-polysaccharide-binding protein and thrombin were identified from trout liver. It is intriguing that complement components such as factor B/C2 and CD59, which are expressed constitutively in mammals, showed remarkable increase in their expression levels in response to the stimuli in bony fish, suggesting that some complement components are acute phase reactant in bony fish.

利用抑制性消减杂交技术克隆了鲤鱼和虹鳟鱼急性期反应物的互补DNA。海藻酸钠、松节油、硬葡聚糖和福尔马林灭活的鳗弧菌被用作刺激剂。结果，从鲤鱼中鉴定了约50个基因，包括哺乳动物中已知的急性期蛋白，如五聚蛋白和血清淀粉样蛋白A，以及几种趋化因子、趋化因子受体、一种新的C型凝集素、一种新的补体因子B同种型、胶质成熟因子、IL-1 β和前B细胞增强因子。另一方面，从鳟鱼肝脏中鉴定了36个基因，包括LECT 2趋化因子、补体B/C2、C型凝集素、纤维蛋白原、结合珠蛋白、血凝素、CD 59、溶菌酶、卵黄蛋白原、前细胞因子、Toll样受体、转铁蛋白、血清淀粉样蛋白A、C-多糖结合蛋白和凝血酶。有趣的是，补体成分如B/C2因子和CD 59在哺乳动物中组成型表达，在硬骨鱼中响应刺激时其表达水平显着增加，表明某些补体成分是硬骨鱼的急性期反应物。

项目成果

K.Fujiki, D.H.Shin, M.Nakao and T.Yano.: "MOLECULAR CLONING AND EXPRESSION ANALYSIS OF CARP(CYPRINUS CARPIO) INTERLEUKIN-1B, HIGH AFFINITY IMMUNOGLOBULIN E FC RECEPTOR GAMMA SUBUNIT AND SERUM AMYLOID A"Fish Shellfish Immunol.. 10. 229-241 (2000)

K.Fujiki、D.H.Shin、M.Nakao 和 T.Yano.：“鲤鱼 (CYPRINUS CARPIO) INTERLEUKIN-1B、高亲和力免疫球蛋白 E FC 受体伽马亚基和血清淀粉样蛋白 A 的分子克隆和表达分析”鱼贝类免疫学

C.J.Bayne, L.Gerwick, K.Fujiki, M.Nakao and T.Yano.: "IMMUNE RELEVANT GENES IDENTIFIED IN THE LIVER OF RAINBOW TROUT, ONCORHYNCHUS MYKISS, BY MEANS OF SUPPRESSION SUBTRACTIVE HYBRIDIZATION."Dev.Comp.Immunol.. 24. 205-217 (2001)

C.J.Bayne、L.Gerwick、K.Fujiki、M.Nakao 和 T.Yano.：“通过抑制消减杂交在虹鳟鱼肝脏中鉴定出免疫相关基因。”Dev.Comp.Immunol..

Fujiki K.,Bayne C.J.,Shin D.-H.,Nakao M.& Yano T.: "Molecular cloning of carp (Cyprinus carpio) C-type lectin and pentraxin by use of suppression subtractive hybridisation"Fish & Shellfish Immunology. 11(In press). (2001)

藤木 K.、Bayne C.J.、Shin D.-H.、Nakao M.

T.Nagai, J.Mutsuro, M.Kimura, Y.Kato, K.Fujiki, T.Yano and M.Nakao: "A NOVEL TRUNCATED ISOFORM OF THE MANNOSE-BINDING LECTIN-ASSOCIATED SERINE PROTEASE (MASP) FROM THE COMMON CARP (CYPRINUS CARPIO)"Immunogenetics.. 51. 193-200 (2000)

T.Nagai、J.Mutsuro、M.Kimura、Y.Kato、K.Fujiki、T.Yano 和 M.Nakao：“一种来自鲤鱼的甘露糖结合凝集素相关丝氨酸蛋白酶 (MASP) 的新型截短亚型

M.Nakao, J.Mutsuro, R.Obo, K.Fujiki, M.Nonaka and T.Yano: "MOLECULAR CLONING AND PROTEIN ANALYSIS OF DIVERGENT FORMS OF THE COMPLEMENT COMPONENT C3 FROM A BONY FISH, THE COMMON CARP(CYPRINUS CARPIO)"European J.Immunol.. 30. 858-866 (2000)

M.Nakao、J.Mutsuro、R.Obo、K.Fujiki、M.Nonaka 和 T.Yano：“来自鲤鱼（鲤鱼）的补体成分 C3 不同形式的分子克隆和蛋白质分析