研究彙報第六十一期 ， Page 1-11，出版時間：1998-12， 詳細內容

利用PCR選殖落花生rDNA之IGS區域

作者：蔡奇助　黃勝忠

摘　　要

　　本研究設計一組引子(primer)，序列分別為IG1: 5' TTGCTGCCACGATCCACTGAG 3'和IG2: 5' CTACTGGCAGGATCAACCAGG 3'。此引子組經聚合酵素連鎖反應(polymerase chain reaction, PCR)，可將各品種的落花生之核糖體DNA (rDNA)基因間隔區(intergenic spacer, IGS)有效選殖(cloning)出來。經電泳分離，其PCR產物長約1,650 bp。將其中一個PCR產物進行部分定序(sequencing)，以進一步確認PCR所選殖出的DNA片段確為rDNA的IGS區域。定序結果與其它植物之相對應區域進行序列比對，証實我們所設計的引子組確能將落花生rDNA的IGS區域選殖出來。

關鍵字： 聚合酵素連鎖反應、核糖體 DNA、基因間隔區、落花生、選殖。

Cloning of Intergenic Spacer Region of rDNA in Peanut by Polymerase Chain Reaction

Chi-Chu Tsai and Sheng-Chung Huang

Summary

　　One set of primers, IG1: 5' TTGCTGCCACGATCCACTGAG 3' and IG2: 5' CTACTGGCAGGATCAACCAGG 3', was designed for cloning the intergenic spacer (IGS) of ribosomal DNA (rDNA) in peanut by using polymerase chain reaction (PCR). The primers could efficiently amplify the IGS of rDNA in peanut. After separating by agarose gel, the length of PCR products was approximately 1,650 bp. The PCR products were then sequencing analyzed and compared with IGS regions of other plant species. The result suggests that it is the IGS region of peanut. This paper present that the PCR techniques could be applied to clone the IGS of rDNA from peanut with a specific set of designed primers.

Keywords: polymerase chain reaction, ribosomal DNA, intergenic spacer, peanut, cloning.