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Published on: November 23, 2016
[Cloning and prokaryotic expression of lipid storage droplet protein 5 gene]
1Department of Pathology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. liqing@fmmu.edu.cn
Aim:
To clone the Lipid storage droplet protein 5 (LSDP5) and express it in E.coli.
Methods:
LSDP5 gene was amplified by PCR from the mouse liver cDNA library, and inserted into pMD18-T vector and sequenced. After analysis of hydrophobicity and secondary structure using software, a fragment of LSDP5 was subcloned into GST fusion expression vector, and the recombinant expression plasmid pEGX-LSDP5 was constructed and expressed in E.coli BL21.
Results:
LSDP5 gene was obtained and its sequence was identical with that reported in GenBank. The E.coli containing pEGX-LSDP5 could express a 40 kD protein after induction of IPTG.
Conclusion:
A confirmed LSDP5 gene is cloned and expressed in E.coli successfully, which lay the foundation for the related research of this gene.
