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[CDNA cloning of human leptin and its expression]
Zhen-Yu Jia1, Xiao-Min Fu, Ai-Hua Jin
1Zhejiang Academy of Medical Sciences, Hangzhou 310013, China. zhenyujia@yahoo.com
Summary
Researchers successfully cloned the human leptin gene cDNA and produced soluble leptin protein in vitro. This advancement supports further investigation into leptin-binding proteins.
Area of Science:
- Molecular Biology
- Protein Expression
Context:
- Leptin is a crucial hormone involved in regulating energy balance and metabolism.
- Understanding leptin's interactions requires a reliable source of functional leptin protein.
Purpose:
- To clone the cDNA of the human leptin gene.
- To express and obtain soluble human leptin protein with a 6x His-tag for future studies.
- To facilitate research on leptin-binding proteins.
Summary:
- Human leptin gene cDNA was cloned using over-hang extension PCR from genomic DNA.
- The cDNA was subcloned into the pIVEX2.3MCS vector for in vitro expression.
- Soluble fusion protein was expressed using the Roche Rapid Translation System (RTS), confirmed by SDS-PAGE and Western blot.
Impact:
- Provides a foundational basis for identifying and studying novel leptin-associated proteins.
- Enables further research into the functional roles of leptin and its interactions.
- Supports the development of new therapeutic targets related to metabolic disorders.