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[Construction of eukaryotic fusion expression vectors of LSDP5 and cellular localization]
Fang Liu1, Li-jun Zhang, Jing Ye
1Department of Pathology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
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.
