摘要
目的:测定重组质粒中编码人蛋白激酶CK2α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CK2α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ABIPRISM310型基因分析仪进行DNA测序。结果:在CK2α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)%。
Objective:In order to screen the recombinant plasmid containing correct insert fragment, cDNAs encoding human protein kinase CK2 α and β subunits in recombinant plasmids were sequenced. Methods: Every four positive clones in recombinant plasmids of human protein kinase CK2 α and β subunits were selected at random. The cDNA inserts were sequenced with the start or end or/and middle region of forward or reverse primers by using dRhodamine Terminator Cycle Sequencing Ready Reaction Kit on ABI PRISM 310 Genetic Analyzer. Results: The results of sequencing showed that two of every four α and β clones possessed correct sequence of encoding human protein kinase CK2 α and β subunits,the two other α and β clones contained at least one mutant base.The average accuracy of 24 sequencing reactions is (98.7±0.4)%,N base average rate is (1.4±0.4)%. Conclusion: Recombinant plasmids containing completely correct sequence of human protein kinase CK2 α and β subunits cDNA were screened successfully by DNA sequencing.
出处
《广东医学院学报》
1998年第4期313-316,共4页
Journal of Guangdong Medical College
基金
广东省卫生厅青年科研基金
粤西高校实验中心建设专款资助