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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | 成之綱 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:14:38Z | - |
| dc.date.available | 2021-07-01T08:14:38Z | - |
| dc.date.issued | 1989 | |
| dc.identifier.citation | Ausubel, F.M. et al. (1987), “Current Protocols in Molecular Biology”, John, Wiley & Sons Inc. pressed.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75681 | - |
| dc.description.abstract | 同位序列(Homeobox)是個具有180個鹼基對的核?酸序列，最初在果蠅(Drosophila melanogaster)中被發現。時至今日，已經知道在酵母菌，線蟲其他昆蟲到脊椎動物如青蛙，老鼠，和人的DNA中都具有如此的序列。大部份具有同位序列的基因，在發生中，扮演重要的角色，而有時間及分佈上的變化。同位序列存在在這些基因當中，主要可轉讀出同位勝?區(Homeodomain), 能夠與DNA作用，也提供吾人瞭解這系列基因功能的可能。 依循著同位序列，在許多物種中存在的事實，吾人尋求同位序列基因在魚體中的存在及表現，以期建立魚的發生模式做為更高等生物的指標。首先，抽取鯉魚(Cyprinus carpia)，斑馬魚(Brachydanio rerio)，吳郭魚(Oreochromis sp.)的基因體DNA與果蠅的同位元模式基因(Homeotic genes)- Antennapedia的同位序列(BamHI/kpnI片段)，以放射性標誌標定後，行南氏雜交反應，以證明魚體中同位序列基因的存在。在各魚經Eco RI限制水解後的基因體DNA中，有約— 4Kb與—>22Kb的DNA片段上有同位序列，顯示魚中至少有兩個Antp—型的同位序列基因的存在，較之其他脊椎動物數目少些． 考慮同位序列基因在鯉魚中的表現，吾人利用北方雜交法，發覺此類基因不只在胚胎發育過程中，而且在成體中皆有表現，因此可能參與發生過程及組織特化的角色。在鯉魚的發生中，主要的轉錄為3.3與1.7Kb RNA以及2.9與1.52Kb RNA，前者從受精卵到原口閉鎖間表現；而後者則是胚體形成到胚胎後期中產生，最強的表現是在體節形成，尾部發生以及心跳期時偵測出。至於在成體中，則以腎，腸，及testis中反應最強；肝，心及眼（包括神經）中次之，顯現出組織特化的功能。 利用分子選殖法，吾人嘗試獲得鯉魚的同位序列基因，並尋求特定基因的結構及表現。所以構築了三種DNA基因庫：一以質體puc 18帶前述4Kb片段；另外則以噬菌體EMBL3帶鯉魚經部份分解的基因體DNA以及用λ gt10帶從鯉魚腎及胚胎中poly A+RNA製造的cDNA。從鯉魚的基因體基因庫中，獲得一個含有同位序列的22kb DNA片段，暫時命名為Chox-X(Carp homeobox-X)。同法篩選cDNA基因庫，獲得四株噬菌體，有強烈的雜交反應，一株來自腎cDNA, 其他來自胚胎。相信再假以時日，吾人即可瞭解鯉魚的同位序列基因的構造，功能，以及發生機制。 | zh_TW |
| dc.description.abstract | The homeobox is a 180 bp DNA sequence characteristically found in Drosophila, however, it was discovered such sequence from yeast, nematode to vertebrate, such as toad, mouse and human. Most of those genes which contain homeobox are expression with temporal and spatial to form special pattern either in embryo or adult. According to the fact that homeobox conserve in different species, we try to search the homeobox genes and their expression in the system of fish. We purified the genomic DNA from carp (Cyprinus carpo), zebrafish (Brachydanio rerio) and tilapia (Oreochromis sp.), and hydrolyzed with the restriction endonuclease of Eco RI and Bam HI; and then, hybridized with the homeobox DNA which was isolated from the gene of Antennapedia in Drosophila. There are two major bands which we acquired in this experiment, one is large than 22kb, and the other 4 kb. We constructed three kinds of DNA liberary: one adopts plasmid of pUC 18 as the vector to ligate with 4 kb fragment which have been described previously, another is the total genomic DNA liberary from carp, and the other are the cDNA liberaries which derived from the carp mRNA of embryo and kidney. After screening, we acquired one homeobox-containing DNA fragment from the total genomic DNA liberary, and named temporarily with chox-x (carp horneobox - x). The transcripts of homeobox genes in carp were identified by Northern hybridization, and showed that their expression are stage and organ dependant in enbryogenesis and adult. The major transcripts during embryogenesis are 3.3, 1.7 Kb RNA and 2.9, 1.52 Kb RNA, the former express from fertilization to blastopore closing; the latter from germ layer formation to hatching, especially, in the stage of somite formation, tail formation, and heart-beat. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:14:38Z (GMT). No. of bitstreams: 0 Previous issue date: 1989 | en |
| dc.language.iso | zh-TW | |
| dc.title | 鯉魚同位序列基因的分子選殖及基因表現 | zh_TW |
| dc.title | Molecular Cloning and Gene Expression of Homeobox Genes in Common Carp (Cyrinus carpio L.) | en |
| dc.date.schoolyear | 77-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 85 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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