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題名 | 體內發育、體外生產與體細胞核轉置牛胚之重要基因表現分析=Gene Expression of in Vivo, in Vitro Produced and Somatic Cell Nuclear Transferred Bovine Embryos |
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作者姓名(中文) | 蕭振文; 蔡麗卿; 曲鳳翔; 劉瑞珍; 劉振發; 李善男; 陳立人; | 書刊名 | 畜產研究 |
卷期 | 38:3 民94.09 |
頁次 | 頁183-195 |
分類號 | 437.33 |
關鍵詞 | 牛胚; 基因表現; 細胞核轉置; Bovine embryo; Gene expression; Somatic cell unclear transfer; |
語文 | 中文(Chinese) |
中文摘要 | 本研究主要目的在探討體內發育(in vivo)、體外受精與體細胞核轉置牛胚於不同發育階段(2細胞期至囊胚期)之重要基因表現。體外生產的牛胚(in vitro production, IVP)、體細胞核轉置(somatic cell nuclear transfer, SCNT)牛胚及田間牛胚非外科採集取得之部分活體牛胚,利用萃取套組純化mRNA後,利用反轉錄-聚合酶連鎖反應(reverse transcription polymerase chain reaction, RT-PCR)獲得互補DNA(complementary DNA, cDNA),再利用合成的引子進行PCR以分析重要基因的表現。結果顯示,分析的19種基因中,對照之常駐(HOUSEKEEPING)基因β-actin與glyceral-dehyde-3-phosphate dehydrogenase (GAPDH)、E-cadherin (E-cad)、octamer-binding transcription factor (Oct-4)、glutamine synthetase (GS)、heat shock protein 70.1 (Hsp)、plakophilin (Plako)與poly (A) polymerase (Poly A)在不同來源、不同胚期的全部牛胚均有表現。因胚源與胚期不同而表現差異的基因有α-2 macroglobulin (A2M)、flavin-containing mono-oxygenase (FMO)、glucose transporter 1(Glut-1)與IGF-I receptor (IGF-Ir)。而在不同胚源、不同胚期均未測到表現的基因transporter 1(Glut-1)與IGF-I receptor (IGF-Ir)。而在不同胚源、不同胚期均未測到表現的基因有connexin43(Cx43)、Cu/Zn superoxide dismutase (Cu/Zn SOD)、desmoglein 1 (Dg1)、IGF-I、interferon tau (IF- )與Mn SOD。為了確認基因之表現是否正確,將測得表現的5種基因產物選殖入pGEN-T Esay Vector system II並進行DNA序列分析,結果與BLAST網路資料庫比對後顯示為正確之序列。本研究之方法與結果可做為牛胚發育生物學與探索基因表現之參考。 |
英文摘要 | The purpose of this study was to compare the gene expression patterns for in vitro produced (IVP), nuclear transferred and in vivo derived embryos at specific developmental stages. The reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the gene expression. In order to further identify these genes, the PCR products of five genes were subcloned and their DNA sequences were determined. Messenger RNA was isolated from bovine embryos produced by IVP, SCNT and in vivo at 2-cell, 4-cell, 8-cell, morula, and blstocyst stages. The cDNA was obtained by RT-PCR and used for gene expression analysis by PCR. Results showed that the gene expression of β-actin, GAPDH, Oct-4, GS, Hsp, Plako, and PolyA were observed in all embryos inspected. The expression of A2M, FMO, Glut-1 or IGF-Ir genes was specific to different embryos sources and developmental stages. The expression of Cx43, Cu/Zn SOD, Dg1, E-cad, IGF-I, IF- and Mn SOD were not detected in all embryos. These results and methods used in this study demonstrated the expression profile of the important genes involved in bovine embryo development. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。