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標題: | 莫三比克吳郭魚性釋素cDNA分子選殖及其在卵巢之表現 GnRH cDNA cloning and its expression in ovary of tilapia ,Oreochromis mossambicus. |
作者: | 郭明偉 |
出版年 : | 1998 |
學位: | 碩士 |
摘要: | 本論文以吳郭魚為實驗對象,利用RT-PCR及5’-RACE的方式初步確定吳郭魚腦部有三種形式的性釋素(gonadotropin-releasing hormone, GnRH),即sGnRH、cGnRH-II及sbGnRH。經序列分析得到134bp長度的sGnRH前驅物之部份序列,包括可轉譯成具功能的sGnRH、酵素辨識的切位(Gly-Lys-Arg)及部份GnRH-associated peptide (GAP)片段;366bp長度的cGnRH–II前驅物之部份序列,包括5’端未轉譯區域(5–UTR),以及可轉譯成signal peptide、具功能的cGnRH-II、酵素辨識的切位與部份GAP片段:216bp sbGnRH前驅物之部份序列,包括5’-UTR,以及可轉譯成signal peptide、具功能的SbGnRH、酵素辨識的切位輿部份GAP片段。此外,另以RT-PCR方法檢視卵巢三種形式之GnRH表現,得到如下結果:若以腦部及卵巢RT-PCR?物大小-樣?判斷依據,則吳郭魚的卵巢有三種形式的GnRH表現,其中以cGnRH-II的相封表現量較多,但也具有少量sGnRH及sbGnRH的表現,雖然如此,但此研究似未能完成銳明吳郭魚卵巢內所?生的三種形式GnRH在卵巢濾胞發育中扮演之角色,因此有待進一步之探討。 In this study, the partial cDNA encoding for the sGnRH precursor, cGnRH-II precursor and sbGnRH precursor were isolated from tilapia brain using RT-PCR and 5’-RACE. The partial 145bp cDNA encoding sGnRH precursor is composed of the biologically active sGnRH, the cleavage site (Gly-Lys-Arg), and a partial GnRH-associated peptide (GAP). The partial 366bp cDNA encoding cGnRH-II precursor is composed of 116bp 5’-untranslation region (5’-UTR), the signal peptide, the biologically active cGnRH-II, the cleavage site, and a partial GAP. The partial 216bp cDNA encoding sbGnRH precursor is composed of 69bp 5’-UTR, the signal peptide, the biologically active sbGnRH, the cleavage site, and a partial GAP. In order to investigate the expression of three forms of GnRH in tilapia ovary, total RNA from ovary of female tilapia at different ovary stages was amplified by RT-PCR. The predicted sizes of RT-PCR products were amplified in ovary. These results demonstrate that three forms of GnRH precursor (sGnRH, cGnRH-II and sbGnRH) mRNA are expressed in tilapia ovary tissue and that high level of cGnRH transcript and low level of sGnRH and sbGnRH transcript are expressed in tilapia ovary. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76338 |
全文授權: | 未授權 |
顯示於系所單位: | 漁業科學研究所 |
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