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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 張繼堯(Chi-Yao Chang) | |
dc.contributor.author | Shih-Jou Tseng | en |
dc.contributor.author | 鄭詩柔 | zh_TW |
dc.date.accessioned | 2021-06-13T00:01:32Z | - |
dc.date.available | 2010-06-07 | |
dc.date.copyright | 2007-07-31 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-30 | |
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Veterinary Immunology and Immunopathology. 95:81-90. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28137 | - |
dc.description.abstract | 石斑魚為亞、熱帶珊瑚礁魚類,是台灣及東南亞地區重要的養殖魚種,肉質鮮美,產量逐年增加,供不應求。由於高密度之養殖方式,造成疾病叢生,為了了解石斑魚的免疫系統以作為防治之基礎,我們進行石斑魚免疫球蛋白M(IgM)的研究。石斑魚IgM是由重鏈與輕鏈組成的四聚體,之前本實驗室已自點帶石斑(Epinephelus coioides)選殖出四型的IgM重鏈基因(Cheng et al., 2006),本研究進一步選殖並分析其輕鏈基因。利用本實驗室選殖出的一段978 bp之IgM輕鏈基因為探針,自石斑魚頭腎cDNA基因庫中選殖出20個全長的石斑魚輕鏈基因。此輕鏈cDNA基因大小介於708至747 bp之間,在5’及3’端具有非轉譯區,其開放讀架區可轉譯為分子量24.6-26.5 kDa之輕鏈。N端leader之後為變異區及保守區。變異區由FR與CDR組成。保守區中含有3個保守性高的cysteine,這些cysteine被認為是輕鏈內部以及其與重鏈之間形成雙硫鍵的橋樑。依據保守區的胺基酸序列,可將這些基因歸納為四種類型,而其變異區則可區分為五種家族。J區連接變異區及保守區,依照J區之N端及C端的序列差異可以推論VJC組合方式具有多樣性,本實驗共找到十種不同的C端序列組合。變異區和保守區與其他已發表的魚類IgM輕鏈基因相比有很高的相似性,依據保守區演化樹分析,我們發現石斑魚具有四種類型IgM輕鏈,比其他魚種的IgM輕鏈類型多。RT-PCR之分析結果,顯示四類型IgM輕鏈在石斑魚頭腎組織中皆有表現。 | zh_TW |
dc.description.abstract | Groupers, coral fish species that inhabit in tropical and subtropical regions, are critical to Taiwanese aquaculture as well as to the rest of Southeast Asia. Due to the culinary value and the ever-growing demand, the yields of groupers progressively increase year after year. However, the highly populated breeding methods within confined or limited habitation space are prone to induction and prevalence of diseases. In order to strengthen the groundwork of disease control and preventative research via a better understanding of grouper's immune system, we initiated this study concerning immunoglobulin M (IgM) of groupers. Immunoglobulin M is a tetramer that consists of two heavy chains and two light chains. Our previous findings identified and cloned 4 CH families from Epinephelus coioides (Cheng et al., 2006). Based on that result, we further analyzed the light chain genes in this proposed study. Utilizing a 1006 bp light chain gene that developed by our laboratory as a probe, we cloned 20 full-length light chains from grouper's head kidney cDNA library. The length of this light chain gene is between 708 to 747 bp, with a non-translated region at 5' and 3' end. In addition, its ORF can be translated into a light chain with molecular weight of 24.6-26.5 kDa. The variable and constant regions are located behind the leader. The variable region is composed of FRs and CDRs; while the constant region consists of 3 highly constant cysteines. It has been suggested that these cysteines formed intradomain and interchain disulfide bonds. According to the amino acid sequences of the constant region, these genes can be classified into 4 isotypes; while the variable region can be distinguished into 5 families. Based on the difference in N- and C-terminal in J segment, J segment can bridge variable and constant region with varieties of VJC combinations, which make it diversity. Overall, we've found 10 different sequences of C-terminal. According to the phylogenetic analysis of grouper's IgM light chains constant region to that of other fish species, we've found that groupers possess 4 isotypes of IgM light chains, relatively higher than other teleost fish. Furthermore, the result of RT-PCR analysis indicated that there are 4 isotypes of IgM light chains expressed in grouper's head kidney tissue. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T00:01:32Z (GMT). No. of bitstreams: 1 ntu-96-R94b45026-1.pdf: 3206538 bytes, checksum: ce9eb215d3990eadcb020333990473a5 (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 口試委員會審定書------------------------------------------I
誌謝-----------------------------------------------------II 中文摘要------------------------------------------------III 英文摘要--------------------------------------------------V 第一章 前言---------------------------------------------1 1.1 石斑魚-------------------------------------------1 1.2 石斑魚常見疾病-----------------------------------2 1.3 魚類免疫-----------------------------------------3 1.4 免疫球蛋白---------------------------------------6 1.5 實驗目的-----------------------------------------9 第二章 材料與方法--------------------------------------10 2.1 石斑魚腎臟cDNA基因庫之構築----------------------10 2.2 石斑魚腎臟cDNA基因庫效價之測定------------------10 2.3 石斑魚IgM gL 1-1質體之大量製備------------------13 2.4 藉EcoRI處理以獲得IgM輕鏈DNA片段-----------------14 2.5 放射性標定石斑魚IgM輕鏈探針之製備---------------15 2.6 石斑魚IgM輕鏈基因之選殖-------------------------16 2.7 質體載體自λ噬菌體載體切出-----------------------19 2.8 小量質體之製備----------------------------------20 2.9 以EcoRI切出IgM輕鏈基因並定序--------------------21 2.10 序列比對及演化樹分析---------------------------22 2.11 頭腎總RNA之抽取--------------------------------22 2.12 設計四種IgM輕鏈專一性之引子--------------------24 2.13 反轉錄-聚合酶鏈反應----------------------------26 第三章 結果--------------------------------------------28 3.1 石斑魚腎臟cDNA基因庫之效價----------------------28 3.2 石斑魚IgM gL 1-1質體之大量製備------------------28 3.3 藉EcoRI處理以獲得IgM輕鏈DNA片段-----------------28 3.4 放射性標定IgM 輕鏈探針之製備--------------------28 3.5 石斑魚IgM輕鏈基因之選殖-------------------------29 3.6 以EcoRI切出IgM輕鏈基因片段並做DNA定序-----------29 3.7 IgM輕鏈保守區-----------------------------------30 3.8 IgM輕鏈變異區-----------------------------------30 3.9 IgM輕鏈之J區分析--------------------------------31 3.10硬骨魚類IgM輕鏈保守區之演化樹------------------------31 3.11石斑魚頭腎組織各類型的IgM輕鏈之表現------------------32 第四章 討論--------------------------------------------33 第五章 參考文獻----------------------------------------36 第六章 圖表--------------------------------------------43 圖一 質體gL 1-1 DNA電泳圖--------------------------------43 圖二 放射性探針強度之測定--------------------------------44 圖三 訊號強弱不同之溶菌斑--------------------------------45 圖四 EcoRI處理各選殖株之電泳圖---------------------------46 圖五 第一類型IgM輕鏈保守區(CL 1)-----------------------47 圖六 第二類型IgM輕鏈保守區(CL 2)-----------------------48 圖七 第三類型IgM輕鏈保守區(CL 3)-----------------------49 圖八 第四類型IgM輕鏈保守區(CL 4)-----------------------49 圖九 石斑魚四種IgM輕鏈保守區胺基酸序列之比對-------------50 圖十 石斑魚IgM輕鏈保守區胺基酸序列之比對-----------------51 圖十一 第一家族IgM輕鏈變異區之胺基酸序列(VL 1)---------52 圖十二 第二家族IgM輕鏈變異區之胺基酸序列(VL 2)---------53 圖十三 第三家族IgM輕鏈變異區之胺基酸序列(VL 3)---------54 圖十四 第四家族IgM輕鏈變異區之胺基酸序列(VL 4)---------55 圖十五 第五家族IgM輕鏈變異區之胺基酸序列(VL 5)---------56 圖十六 石斑魚五種IgM輕鏈變異區之胺基酸序列之比對---------57 圖十七 石斑魚IgM輕鏈中的J區胺基酸序列--------------------58 圖十八 石斑魚與其他魚種IgM輕鏈保守區之演化樹分析---------59 圖十九 選殖株19-1保守區之核苷酸序列----------------------60 圖二十 選殖株10-6保守區之核苷酸序列----------------------61 圖二十一 選殖株19-6保守區之核苷酸序列--------------------62 圖二十二 選殖株19-22保守區之核苷酸序列-------------------63 圖二十三 石斑魚IgM輕鏈四類型引子專一性之測定-------------64 圖二十四 石斑魚頭腎組織各類型的輕鏈IgM之表現-------------65 表一 石斑魚四種IgM輕鏈之保守區(CL)相似度---------------66 表二 石斑魚五種IgM輕鏈之變異區(VL)相似度---------------67 | |
dc.language.iso | zh-TW | |
dc.title | 石斑魚免疫球蛋白M基因之選殖及分子特性 | zh_TW |
dc.title | Cloning and Molecular Characterization of Grouper Immunoglobulin M (IgM) Genes | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭應誠(Ing-Cherng Guo),林富邦(Fu-Pang Lin) | |
dc.subject.keyword | 點帶石斑,石斑魚,免疫球蛋白,輕鏈, | zh_TW |
dc.subject.keyword | Epinephelus coioides,Grouper,Immunoglobulin,IgM,Light chain, | en |
dc.relation.page | 67 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-31 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
顯示於系所單位: | 漁業科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-96-1.pdf 目前未授權公開取用 | 3.13 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。