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頁籤選單縮合
| 題 名 | Oct-4基因於體外受精與體細胞核轉置牛胚之表現=The Oct-4 Gene Expression at Different Stages of Bovine Embryos Produced by in Vitro and Nuclear Transfer |
|---|---|
| 作 者 | 蕭振文; 蔡麗卿; 陳裕信; 曲鳳翔; 楊鎮榮; 李善男; 陳立人; | 書刊名 | 畜產研究 |
| 卷 期 | 37:4 2004.12[民93.12] |
| 頁 次 | 頁301-311 |
| 分類號 | 437.3 |
| 關鍵詞 | 牛胚; 基因分析; 發育; Bovine embryo; Gene analysis; Development; Octamer-binding transcription factor 4; Oct-4; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 本研究之主要目的,乃應用反轉錄-聚合酶連鎖反應(reverse transcription-polymerase chain reaction, RT-PCR)分析以不同方式獲得的牛胚於不同發育階段Octamer-binding transcription factor 4 (Oct-4)與β-actin mRNA之表現型式。同時,將此二種基因之PCR產物選殖並分析DNA序列,以確認Oct-4與β-actin二種基因之正確性。分析之牛胚-是利用體外生產系統(in vitro production, IVP)與核轉置(nucleus transfer, NT)方法生產的2細胞期、4細胞期、8細胞期、桑椹期及囊胚期牛胚;二是田間牛胚非外科採集所取得之體內桑椹期及囊胚期牛胚,以套組萃取不同胚期牛胚之mRNA,進行RT-PCR獲得互補DNA(complementary DNA, cDNA),再利用分析Oct-4與β-actin基因的引子對進行PCR。試驗結果顯示,利用IVP與NT所生產的2細胞期、4細胞期、8細胞期、桑椹期及囊胚期牛胚,以及採集自牛隻體內桑椹期及囊胚期的牛胚樣品,均有Oct-4與β-actin之mRNA表現。而Oct-4與β-actin基因經過選殖與DNA序列分析後,進入NCBI進行DNA序列比對,結果Oct-4基因與Bos taurus者完全相符。而做為對照的β-actin基因亦正確表現,顯示在牛胚不同發育階段,不論是IVP、NT或體內胚,均可以見到此兩種基因的mRNA表現。本研究結果可供開發檢測牛胚發育潛力之分子標記之用。 |
| 英文摘要 | The purpose of this study was to investigate the expression of Oct-4 and housekeeping gene βactin in specific developmental stages of bovine embryos. Those embryos were produced by in vitro production (IVP) and uncleus transfer (NT), and developed in vivo. The reverse transcription-polymerase chian reaction (RT-PCR) was used to measure the gene expression in order to further identify these genes’ amplification, the PCR products specific to Oct-4 and β-actin were subcloned and DNA sequences were determined. Messenger RNA was isolated from bovine embryos produced by IVP, NT and in vivo at 2-cell 4-cell, 8-cell, morula, and blstocyst stage. The cDNA was obtained by RT-PCR and used as a source for PCR analysis of Oct-4 and β-actin gene expression. Results showed that the expression of Oct-4 andβ-actin housekeeping gene could be detected at different stages of embryos from IVP, NT and in vivo. The results demonstrated that RT-PCR technique was a valuable tool for the analysis of important genes in bovine embryos. This system could be used in the development of molecular markers for early screening and quality determination of bovine embryos during the early stage of development. |
本系統中英文摘要資訊取自各篇刊載內容。