[Construction of eukaryotic fusion expression vectors of LSDP5 and cellular localization].

Aim: To construct the eukaryotic fusion expression vectors and localization of (lipid storage droplet protein 5, LSDP5) in COS7 cells.

Methods: Eukaryotic expression plasmid pCMV5-HA was digested with Nde I and BamH I and pMD18-LSDP5 plasmid was digested with Bgl II and Nde I. Full length LSDP5 was subcloned into pCMV5-HA. After confirmed by restriction enzyme digestion analysis and sequencing, the plasmid pCMV5-HA-LSDP5 was transfected into COS7 cells using Lipofectamine 2000 and the expression and localization of the fusion protein was detected by fluorescent microscope.

Results: Restriction digestions and sequencing assays showed that the recombinant plasmid of pCMV5-HA-LSDP5 was successfully constructed and the fusion proteins were detected in cytoplasm and were targeted to the surfaces of lipid droplets visualized by Bodipy 493/503 in COS7 cells.

Conclusion: The recombined plasmid pCMV5-HA-LSDP5 expressing the fusion protein of LSDP5 and HA-tagged is successfully constructed and the fusion protein distributes in cytoplasm and is co-localized with lipid droplets.