摘要
本文建立了一种根据碱基序列分离和分析DNA片段的新方法,即在含均一浓度变性剂的凝胶电泳过程中形成温度梯度,使分子长度一致而碱基序列不同的DNA片段相互分离。该技术有望在多方面得以应用。
A new technique, named temperature gradient gel electrophoresis (TGGE) ,used for sequence dependent separation of DNA fragments has been devloped. If doublehelical DNA is partially meltiog in polyacrylamide gels, its electrophoretic mobility undergoes a sharp transition, resulting in a large reduction in mobility. In the present experiment, the transition that was effected at a uniform concentration of denaturant, formamide, in a temperature gradient formed during electrophoresis. Each restrictive fragment of pBR322-DNA and bacteriaphage λ-DNA exhibited the mobility transition at a particular temperature. The sudden retardation of fragments moving forward in the gle depended upon nucleotide composition and sequence, rather than the length. When combined with length dependent electrophoresis in the perpendicular direction, TGGE provided a two dimentional separation of DNA restrictive fragments. The resolving power of the new system was demonstrated by the clear resolution of over 80 fragments of the restrictive enzyme Hin fI digest of λ-DNA. This technique would find its application in many fields.
出处
《遗传与疾病》
CAS
CSCD
北大核心
1989年第1期1-3,63+66,共3页
基金
国家自然科学基金
