Construction of the Eukaryotic Expression Vector of 14-3-3

Objective To clone human 14-3-3 gene and construct the eukaryotic expression vector of pcDNA3.1(+)-14-3-3.Methods Total RNA was isolated from human brain tissue.The full-length human14-3-3 cDNA was obtained by RT-PCR and then inserted into pCUm-T vector for sequencing.The correct gene was subcloned into pcDNA3.1(+) to generate recombinant eukaryotic expression vector pcDNA3.1(+)-14-3-3.Results Enzyme digestion analysis PCR and sequencing showed that the target gene was cloned into recombinant vector.Conclusion The eukaryotic expression vector of pcDNA3.1(+)-14-3-3 was constructed successfully,which lay the foundation for the further study of 14-3-3 gene in Parkinson disease.