摘要 目的：测定重组质粒中编码人蛋白激酶CK2 α和β亚基cDNA序列，筛选含完全正确插入片段的重组质粒。方法： 随机选择人蛋白激酶CK2 α和β重组质粒各四个阳性克隆。使用二氯罗丹明荧光标记终止底物循环测序试剂盒，分别加入首尾正反或/和中段正反引物，运用ABI PRISM 310型基因分析仪进行DNA测序。结果：在CK2 α和β阳性克隆中各有两个含完全正确的插入片段的重组质粒，其他各两个重组质粒的插入片段中均存在至少一个碱基突变。24个测序反应的平均精确度为(98.7±0.4)%；N平均出现率为(1.4±0.4)%。结论：通过DNA测序筛选到了含完全正确的人蛋白激酶CK2 α和β亚基cDNA序列的重组质粒。

    关键词 序列分析，DNA；蛋白激酶CK2，人；重组DNA

    Sequencing of cDNAs encoding human protein kinase CK2 α and β subunits in recombinant pkasmids with ABI PRISM 310 Genetic Anakyzer

    Liu Xinguang，Liu Wen，Liang Nianci，et ak

    (Institute of Medicak Biochemistry，Guangdong Medicak Cokkege，Zhanjiang 524023)

    Abstract Objective：In order to screen the recombinant pkasmid containing correct insert fragment， cDNAs encoding human protein kinase CK2 α and β subunits in recombinant pkasmids were sequenced. Methods： Every four positive ckones in recombinant pkasmids of human protein kinase CK2 α and β subunits were sekected at random. The cDNA inserts were sequenced with the start or end or/and middke region of forward or reverse primers by using dRhodamine Terminator Cycke Sequencing Ready Reaction Kit on ABI PRISM 310 Genetic Anakyzer. Resukts： The resukts of sequencing showed that two of every four α and β ckones possessed correct sequence of encoding human protein kinase CK2 α and β subunits ......