變性梯度膠體電泳分析( DGGE; Denaturing Gradient Gel Electrophoresis )在遺傳上的應用

Abstract

變性梯度膠體電詠分析(DGGE；Denaturing Gradient Gel Electrophoresis)，是以DNA的變性原理為基礎而偵測DNA變異的一種生物技術。DGGE偵測DNA的變異有高度靈敏及精確性，因此在人類遺傳疾病的偵測上效果優越，所以目前為止以動物及醫學研究應用DGGE技術較多，植物方面僅有小麥及玉米有這方面的報導。在作物的育種工作中常因DNA標誌多型性不易取得，而使育種工作不易進行。RFLP及RAPD雖然已大量的應用於作物的育種工作中，但對自花授粉的作物仍有不易取得品種間的多型性的缺點。DGGE除了能補救該缺點外，更具有可能發展到一次分析大量樣品的優點，因此應是相當具有潛力的一種新興生物技術。Denaturing gradient gel electrophoresis (DGGE) is a relatively new biotechnological tool which allows the identification of polymorphism based on changes in the melting behavior of DNA fragments. It is highly sensitive and accurate for the resolution of single-base differences along the entire length of the fragments. It has been successfully used as diagonstic tools for detecting heritable human dieseases and medical studues and to construct molecular marker in wheat and maize. DGGE is particularly useful for autogamous crops where intraspecific polymorphisms are rare and difficult to detect by either RFLP or RAPD system. It can combine with RAPD to create polymorphic molecular markers among closely related varieties in high frequency. In addition, with some modifications, DGGE can be transformed into two dimensuonal electrophoretic system which makes it possible to analyze multiple samples in a single assay. These two features makes DGGE as a potential tool for genetic and breeding analysis