摘要
目的：构建带有Ｅ－ｔａｇ蛋白标签的ｐＴＣ０１Ｅ载体．方法：从噬菌粒载体ｐＣＡＮＴＡＢ５Ｅ中ＰＣＲ扩增出Ｅ－ｔａｇ基因片段，经Ｋｌｅｎｏｗ片段补平和ＳａｃＩ消化后，将含有Ｅ－ｔａｇ序列的３８１ｂｐ片段克隆至分泌型原核表达载体ｐＴＣ０１的ＳａｃＩ－ＳａｍＩ位点中．结果：构建成带有Ｅ－ｔａｇ蛋白标签的ｐＴＣ０１Ｅ载体，限制性内切酶图谱和ＤＮＡ序列分析表明构建过程正确．结论：用ＰＣＲ－克隆策略获得了ＤＮＡ序列正确的ｐＴＣ０１Ｅ原核表达载体，它将使外源基因的克隆、表达和检测更加方便、快捷．
Aim: To construct prokaryotic vector pTC01E with E tag segment. Methods: E tag gene fragment was amplified from phagemid pCANTAB 5E, filled by DNA polymerase I Klenow fragment and digested with restriction enzyme Sac I Sam I site of pTC01 plasmid vector to construct the pTC01Ean E.coli expression vector with peptide tag. Results: The results of restriction analysis and DNA sequencing showed that PCR and cloning procedures were successful. Conclusion: Prokaryotic vector pTC01E was acquired successfully by PCR cloning, it will enable the cloning, expression and detection of foreign gene more conveniently.
出处
《第四军医大学学报》
1998年第3期274-276,共3页
Journal of the Fourth Military Medical University
