恶性疟疾重组质粒pPF_（14）DNA探针的制备

The Preparation of Recombinant Clone pPF_(14)DNA Probes

采用基因克隆技术，将恶性疟原虫（Ｐｌａｓｍｏｄｉｕｍｆａｌｃｉｐａｒｕｍ）ｐＰＦ１４质粒转化大肠杆菌ＨＢ１０１，经扩增后碱裂解法提取质粒ＤＮＡ，核酸内切酶酶切图谱分析鉴定后，低熔点胶法回收而获得高纯度和高回收率的插入片段（ｐＰＦ１４ＤＮＡ），经３２Ｐ标记后具较高敏感性与特异性，该探针制备方法具简便、稳定、省时的优点。本文对各制备程序进行了探讨。

This article described the praparation of pPF14 DNA probes. These procedures involved transformation of Escherichia coli strain HB 101, Amplification, plasmid was isolated by alkaline method and was dounbly digested with the restriction enzymes Hind Ⅲ and SaⅠ, pPF14 DNA fragment afterwards was analyzed by agarose gel electrophoresis and low melting gel methed recovered. The results showed that high purity and efficient pPF14 DNA fragment was obtained when using radiolabelled. The probe had a high sensitivity and speciality. The method was much easer,faster, stable and economical than other means, mean while, we discussed these procedures from defferent angles.