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標題: | 鯉魚核醣體蛋白質S17及L37與轉譯因數EFlα之選殖及其在卵發育過程中表現之研究 Molecular Cloning Of The Ribosomal Protein, S17 And L37, And The Elongation Factor lα Of Carp And Their Expression During Oocyte Development |
作者: | Ts'ai-Lien Liao 廖彩蓮 |
出版年 : | 1999 |
學位: | 碩士 |
摘要: | 本實驗是從鯉魚卵巢cDNA庫篩選出轉譯延伸因數1α(EF-lα)與核醣體蛋白(Ribosomal protein L37及S17)的 cDNA。EF-1α cDNA全長1651 bp,轉譯之胺基酸共458個,stop codon位於第1374個核甘酸,3'UTR長247 bp。L37 cDNA全長389 bp,轉譯之胺基酸共92個,3'UTR長100 bp。S17 cDNA全長434 bp,轉譯之胺基酸共135個,3'UTR長 25 bp。它們與其他物種的同源基因相似性相當高。本論文即以此cDNA進行EF-lα、L37及S17在鯉魚卵發育過程中,表現情形的研究,以瞭解鯉魚卵發育過程中,轉譯機構建立的情形。 北方轉印分析(Northern blotting)結果顯示鯉魚卵之EF-lα、L37及S17 mRNA在發育過程的時期Ⅱ濃度最高,但隨著卵成熟而下降,此結果與蛙卵之EF-1α與核醣體蛋白之變化情形相同。另外,西方轉印分析(Western blotting)顯示EF-1α蛋白質在鯉魚卵發育過程累積,於時期Ⅱ濃度最高,至卵成熟時EF-lα含量下降。但本實驗以L37及S17之重組蛋白質所引發的抗體,無法辨認卵內的L37及S17, 故無法瞭解這兩個蛋白質在卵發育過程之變化情形。 Carp elongation factor 1α (EF-1α) and ribosomal proteins(L37 and S17) were isolated from an ovary cDNA library. EF-1α is a 1651bp cDNA and encodes a 458 residue polypeptide. L37 is a 389bp cDNA and encodes 92 residue polypeptide. S17 is a 434bp cDNA and encodes 135 residue polypeptide. They are extremely high homologous to their counterparts in other animals. In this paper, I study the expression change of EF-1α, L37 and S17 during the oogenesis to understand the translation during development of carp oocyte. Northern blotting revealed that EF-1α, L37 and S17 mRNA reach the highest level in the second stage of oogenesis, but they decrease with the oocyte maturation. Western blot analysis indicated that EF-1α protein is accumulated until the second stage and then decreases. However, it failed to detect L37 and S17 proteins in oocytes by antiserum against bacterial recombinant L37 and S17 proteins. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75070 |
全文授權: | 未授權 |
顯示於系所單位: | 動物學研究所 |
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