Construction and identification of eukaryotie expression vector of hsa-miR-200

Objective: To construct and identify the eukaryotic expression vector pcDNA3.1(+)-miR-200a of hsa-miR-200a.It may lay a foundation for further studying the function of hsa-miR-200a.Methods:Amplified by reverse transcription-polymerase chain reaction(RT-PCR),The pre-miR-200a sequence was inserted into pcDNA3.1(+) vector.The recombinant plasmid of pcDNA3.1(+)-miR-200a was confirmed by restriction endnuclease analysis as well as DNA sequencing.Results:The result of DNA sequencing demonstrated that eukaryotic vector pcDNA3.1 (+)-miR-200a was completely correct.Conclusion: The successful construction of eukaryotic expression vector of hsa-miR-200a provided a useful tool for the study of biological function of hsa-miR-200a and its potential target genes.