毛细管电泳单链构象多态性分析检测质粒中基因突变

Detection of vector gene mutation by single strand conformation polymorphism with capillary electrophoresis

目的建立一种检测基因点突变的方法。基因的点突变在癌症的发生中起重要的作用,检测基冈点突变成为分析癌症发生的重要手段。方法采用以一对引物PCR法扩增出的pBR322和pUC19c氨苄抗性基因为点基因突变模型,建立了一种简便快速的毛细管电泳单链构象多态性分析方法,用于分离基因单点突变模型。结果以6％线性聚丙烯酰胺为分离介质,分离温度为19℃,分离电压为13 kV,可分离出模型中的4个单链DNA构象。结论建立的模型是可靠的,分析方法具有快速、灵敏度高的优点。

Objective Gene point mutation plays an important role in cancer occurrence. Detection of gene point mutation is an important method on analysis of cancer occurrence. The purpose was to establish a method to detect gene point mutation. MeThods A pair of primers was used to amplify ampicillin resistance gene of vectors as pBR322 and pUC19c. Using these PCR products as a point mutation gene model developed a single strand conformation polymorphism with laser-induced fluorescence capillary electrophoresis method for detection of point mutation gene model. Results We separated four single strand conformations of the model under conditions of 6% SLPA, 19 ℃ running temperature, and 13 kV running voltage. Conclusion The result revealed that this model is reliable and the method is rapid with high sensitivity.