一种快速简便可靠的穿梭质粒靶基因序列测定方法

A Rapid Simple and Reliable Method for Sequencing the Targeted Gene of Shuttle Vector pSP189

在对突变的分子机理所进行的研究中，ｐＳＰ１８９穿梭载体是一种非常重要且极具使用价值的工具。基于该载体系统，我们探索并建立了一简单、快速和高效的程序步骤用以对载体上的ＳｕｐＦＴＲＮＡ靶基因序列进行测定和分析。在此方法中，我们采用碱变性法对双链质粒ＤＮＡ模板进行变性，再使用５＇端标记有γ－３２Ｐ－ＡＴＰ的引物对所制备的ＤＮＡ单链序列进行测定并获得了满意的实验结果。

The authors established a simple, rapid and effective procedures for sequencing the targeted agene SupF TRNA of shuttle vector pSP189, which is very important and useful in molecular mechnism research of mutagens. In this mehtod, double stranded plasmid DNA as template, prepared single stranded DNA with alkali denaturation, applied r 32P ATP for labelling 5' end of primer, were used.A satisfactory result was achieved.