人核糖体蛋白L6编码基因的克隆及正反义真核表达载体的构建

Cloning of Ribosomal protein L6-encoding gene and constructing of sense and antisense eukaryotic expression vectors

目的:通过克隆人核糖体蛋白L6(RPL6)cDNA序列,构建其正反义真核表达载体,以探讨RPL6与白血病多药耐药的关系。方法:用RT-PCR技术扩增RPL6 cDNA序列,DNA重组技术构建其正反义真核表达载体。结果:成功扩增RPL6 cDNA序列,经DNA测序,证实与GenBank中的人RPL6编码区序列一致,酶切证实目的基因分别按正反两个方向亚克隆至真核表达载体。结论:成功克隆人RPL6 cDNA序列,并构建了正反义真核表达载体。

Objective： To explore the relationship between ribosomal protein L6 and MDR of K562/A02 cell by cloning of RPL6-encoding gene and constructing of sense and antisense eukaryotic expression vectors. Methods： RPL6 cDNA was obtained from its mRNA and amplified by RT-PCR. Results：The cDNA encoding RPL6 from K562/A02 cell was successfully amplified and PCR product was sequenced and identified to be consistent with RPL6 cDNA reported in GenBank. Recombination plasmids containing both sense and antisense RPL6 cDNA were identified by gel electrophoresis analysis using restriction endonuclease. Conclusion： RPL6-encoding gene has been successfully cloned and sense and antisense eukaryotic expression vectors constructed.