Burmeister M, diSibio G, Cox D R, Myers R M
Department of Physiology, University of California, San Francisco 94143.
Nucleic Acids Res. 1991 Apr 11;19(7):1475-81. doi: 10.1093/nar/19.7.1475.
Genomic Denaturing Gradient Gel Electrophoresis (gDGGE) provides an alternative to the standard method of restriction fragment length polymorphism (RFLP) analysis for identifying polymorphic sequence variation in genomic DNA. For gDGGE, genomic DNA is cleaved by restriction enzymes, separated in a polyacrylamide gel containing a gradient of DNA denaturants, and then transferred by electroblotting to nylon membranes. Unlike other applications of DGGE, gDGGE is not limited by the size of the probe and does not require probe sequence information. gDGGE can be used in conjunction with any unique DNA probe. Here we use gDGGE with probes from the proximal region of the long arm of human chromosome 21 to identify polymorphic DNA sequence variation in this segment of the chromosome. Our screening panel consisted of DNA from nine individuals, which was cleaved with five restriction enzymes and submitted to electrophoresis in two denaturing gradient conditions. We detected at least one potential polymorphism for nine of eleven probes that were tested. Two polymorphisms, one at D21S4 and one at D21S90, were characterized in detail. Our study demonstrates that gDGGE is a fast and efficient method for identifying polymorphisms that are useful for genetic linkage analysis.
基因组变性梯度凝胶电泳（gDGGE）为识别基因组DNA中的多态性序列变异提供了一种替代标准限制性片段长度多态性（RFLP）分析方法的技术。对于gDGGE，基因组DNA用限制性内切酶切割，在含有DNA变性剂梯度的聚丙烯酰胺凝胶中分离，然后通过电印迹转移到尼龙膜上。与DGGE的其他应用不同，gDGGE不受探针大小的限制，也不需要探针序列信息。gDGGE可与任何独特的DNA探针联合使用。在这里，我们使用来自人类21号染色体长臂近端区域的探针进行gDGGE，以识别该染色体片段中的多态性DNA序列变异。我们的筛选小组由来自9个人的DNA组成，用5种限制性内切酶切割，并在两种变性梯度条件下进行电泳。我们在测试的11个探针中的9个中检测到至少一种潜在的多态性。详细鉴定了两种多态性，一种在D21S4，另一种在D21S90。我们的研究表明，gDGGE是一种快速有效的识别多态性的方法，这些多态性对遗传连锁分析很有用。
